That's a real overture going
to Russia in about six years.

And yes we're out

there with her pretty careful and

not nothing more than a rock for us.

Thank you very much for inviting me.

And it's a great pleasure to meet
presentation in this building.

I think it's a wonderful
set of events and.

Ironically I for the time I've been here
for six years is my first time when I

have the pleasure to make a presentation
list audience in this auditorium.

My title of my talk because he is
pretty alone and I can essentially

hear because he could summarize
a service I'm going to talk about.

So you can see that I'm we're focused
on studies of chromosome stability and

we carry out our research can use the same
time we think that this study same Porton

because we can expect probably this
to dissolve through our problems

that people deal with a stunt human
to use and human pretty monotheisms.

This building because you know it contains
a lot of labs to doing different research

and recently have been invited
to be on his committees for

several people who are doing
absolutely different things.

Such a study born formation one that.

Aeration.

Obviously the studies are very
far away from what I'm doing.

If you look at the start of the same time.

After being in the communities
where it was understood that

there are some similarities.

Even.

Some approximation were used in similar
terms and started similar problems.

So if you.

Imagine that.

If you come to the statement
that comes from some bones and

if depending how you
draw the even look alike.

So I realize the people who are starred
in how bones are broken and

how they repaired.

They encounter the same questions that
were trying to unravel in the study

chromosome breakage.

First task is need to repair breaks

in both cases and this breaks the must
be repaired pretty accurately.

Otherwise.

If you don't repair the bones
that can lead to lethality or

if you're a parent and accurately it
can lead to a person with disabilities.

The same applies to chromosome breakage.

So there are in some senses
a scaffold of from genetic term

because they carry a lot of our
genes essential genetic information.

So if break is not repaired.

It leads to solve a little it
is solved us or if you heard

him correctly it leads to disabilities
that are many first bite here or

did you see some cancers in the in humans.

The questions that are addressed and
the questions I encounter

Why given this is almost
the same what people are asking

when they study chromosome breaks
why are bones of chromosomes break.

What genetic environmental factors
to dispose them from breakage

how to repair the brakes.

If you simply.

What goes wrong on
a repair is an accurate.

This is so conspiratorial type of
human thought of normal thousand each.

Because by specific color.

Those are very precise carriers
of genetic information and

there are a lot of mechanisms in
South exist that guarantee that.

The chromosomes going to be stable and
interact.

But sometimes something goes wrong and
if you look at

some cancer cell lines you can find the
chromosomes of the fused to each other.

So if you see two colors.

Next to Charlotte means a thirty year
range with most problems translocation

are as a matter fact much jurist of
cancer cell lines characterized by.

Structural all the normal variations in

why it's all happening.

Our problem on call them
which is that can drive our

chromosomes that we also break is
the most dangerous type of damage.

Because one unrepaired break.

Drastically changed the level of
expression of genes for example if.

The broken fragment would be not

repaired to last you can lose
the wall a little that covers.

A mutant another chromosome the process
that the scope of most of you.

There is as I go city that can
lead the development of cancer.

The same time a new pair of
break can be on the selection.

It can be amplified because it
doesn't contain center near and

now it can last a cassock processes
can accumulate in the south and

that leads to example here or
in the transcription factor the drug.

The for the cancer development
there are often the extra

chromosome Lampley fight regions look as
inverted diamonds they call double mine.

It's.

Another consequence of the break that this
broken fragment becomes very prone for

invasions they can invade normal scrum and
so on and

it can lead to transfer occasionally on
and one of the most famous examples is

the formation of the C R A B O fusion
Kylie's that is expressed in chronic.

Neurogenic Meares is the only successful
drug on the market right now that

partly treats the disease.

At the same time.

This can lead to smaller scale variations
that became very popular recently.

Apparently human population
is loaded with so

called copy a number of
variations means that.

Certain genes there are either.

If you look at different individuals
we can cover more of some genes

a loss of some genes even a different
variations in this copy number

of course they contribute to our diseases
but the same time the country would

majorly to the why we look
different from each other.

So what can the agents break chromosomes
what agents can break forceful diasters

morons and allow There are a lot of them
are very well characterized and there

are a lot of there's no humans factor so
we're in the origin starters that.

Break the knee the rocky or
interactive for example and

eyes of radiation and assumption is and
for a long time people believe the last.

Probably the most prominent source of
the damage decisions that can break.

Beneath any given position.

Essentially any religion can be wrong for
this bridge.

Except for the case of good grapes
the interviews by site specific and

the nucleus.

But are the agents there more or less.

The like the specificity.

Never were us in the movie lost
the case became clear the breaks

don't happen randomly and certain
regions are more prone than others.

And people see a thong are the pollution
of the scale or they can see it when

they analyzed chromosomes from
patients that are from cancer or

from here it is useless
if you look closely when

you look at the regions that
are predisposed to breaks

you can find that very often
they contain repetitive if you

know lies gross derangement breakpoint
database or human gene mutation database.

That's the third Those are reports
various of all information.

From sperm or somatic cells translocation

events you very often you can find
repetitive elements over the repeats.

But not only simple repeats but

rather a lot of pieces you
find repeals that can top.

So-called normal but at the information
Google talk about that later.

Are more specific examples of Fragile X.

syndrome that they discovered by Steve
Warren so that results from expensive

E.P.O. and the smell of face chromosome X.
chromosome and you can see that.

Tiny links here that very
often break under cover sleep.

They actually break.

And this is the source.

In the river.

In life because it leads to so
you are absolutely syndrome.

So this may also characterized
by a very severe deficiency is

the frequency of carriers that
carry so-called pretty mutations

that sixty precludes one out of two

hundred women in human population they're
a carrier of this so called premeditation.

I think some students who
are to try to speak up.

That's not not the only place and
you know not only chromosome.

But other positions across
the genome were expansion of the G.

and H. sequences can happen and
predispositions to fragility and

another example is translocation
between chromosome fourteen and

eighteen and that is very common
translocation follicle inform us.

It's caused by break it should be zero two
which contains three picks form in the.

Last but.

Definitely not least
example is the current

translocation stimulated by bracket
palindromic size that translocation

between chromosome eleven and twenty
two one to pull in drums are joined.

To each other.

Usually doesn't cause disease but
it leads to human infertility.

So for so far there are seven
eighty DISH problem drums and

into fourteen human genome.

There are scattered around the genome.

However.

It's known that it's very
difficult to sequence.

How many of this unstable sequences
exist in the human genome.

It's still a big mystery.

This conclusion from this.

He says is that this repeal
can occur the normal.

Or we can hear them from our parents and

then look what is common
feature of the signals is that

implicated in causing breakage causing
rearrangements the common feature

is that although they can adopt
so-called non-canonical Dini structures

such as hair constructions of cruciform
structures people X. or quadruplex the.

Those are different confirmations
that differ from canonical.

How this breaks how the structures.

Lead to break the still
mysterious process.

And if you modify these questions
that were addressed to study.

Borne damage chromosome damage
where the couple more so

in our studies we want to understand
how potent this second destruction

in the Newson chromosome breaks.

Since that is that we can
find that one out of millions

every cell is going to
experience this break.

If for the cell contains
this repetitive sequence.

Y. chromosomes break.

So why architectural change in our D.N.A.

organization can cause
such drastic consequences.

Why chromosomes break.

What factors mediated.

And obviously.

What structural parameters of the piece
make them prone for chromosome break which

not all the things that you can look
analyze are going to cause some breaks.

This is questions we'll try and

address in our work in our studies in
model up with stunning sequence might use.

Formed by pollen drums.

So this is cruciform structure so inverted
is they can adopt cruciform or structures

recently incorporation with
Skeeter out from Emory University.

Studying he did you teach.

I decided to when I thought about
going as a stalker I decided not to

talk about all of this repeats because
otherwise we're going to overwhelm and

I'm going to take an example and
just to show you.

The way the law is
the the breakage that is

form that the cruciform
adoption sequences and

also what are the with of
those you use in what.

And the law is.

The consequence of this breaks and
the mechanism of the breakage.

But essentially the same approaches
we're using to analyze breaks that

happening at this motives as well and

just to be a kind of me if you look
at the mechanism of breakage for

each I repeat for each structure
the mechanism is different and

this is the leaf particular
signature how breaks are happening.

And if you analyze the results and
rearrange means you can play it back and

did use what was causing the factor for
the instability.

OK we're using normal not working with
humans using in the model organism.

So far at least in the near Reaper it's
known that whatever you discover in

the small organism states the same place
through for our mammalian genomes.

At least in Europe or
the way how we detect

breaks we had to build the system and we
wanted to do system which is going to be.

Very easy and also a very very sensitive
to detect chromosome a bridge so

what you see here is the diagram of
particular chromosome chromosome four

the circle means centromere this is
the left arm not the scale of this

chromosome and
first person who not noticed

that there are some some particular
about this chromosome a solution.

Found that there's a natural present
one gene on this chromosome and

the beauty of this location is that if
you go to the left side from Can one.

It contains it doesn't contain any
essential genes which means that

if we break happens in this region.

East.

They're going to survive and
can one is contra select will markers.

It's like a drug.

So I'm not going to go
into these genetics but

the sensual if we select a new
media containing an hour and.

We can select for events one
chromosome is going to be broken and

can one still be lost and

we can money toward using additional
markers in nicely turn the color.

So the usual white they become red.

And.

Here's our insertion side we can
insert any sequence might give and

to analyze how frequently this chromosome
chromosome arm is going to be lost.

So essentially it's going to look
something like this spontaneous rate of.

Armloads is extremely
rare is extremely low.

It's want to.

Five to ten to nine service so

in micro biology terms means
that you need to please to

replicate plate twenty plates and
maybe you're going to find one revert.

It's a very very rare event.

In cartoon way we're looking for
something like that.

So essentially the arm is lost.

And then we're going to look at
this chromosome is going to repair

this broken arm also work
of to do small markers.

As a free one.

Called.

Gene dosage markers usually used
there will not grow on the media

containing copper and here we cover one
gene which is includes the protein.

That scope or but

they can grow on the media with
high concentration of corporate.

If happens.

Essentially we're going to look for this
broken fragment is going to be defied.

And again spontaneous.

The rate of cation is very extremely low.

So as opposed to the we're starting to
pull in sequences or inverted repeats.

In other words in a linguistic way.

Those are very funny sequences really the
same from left to right or right to left.

This is the all the problem
drum it's written

on the fountain in the fear
temple in Constantinople.

And so but with a single it's going to be
the same sentence from left to right and

right to left in English.

If you translate it to school it
was you seems not just you faced

before you enter the church.

You know.

Seems to be in a pool there it's here so

it's going to be the same sequence
that it is from five to three prime

direction opposite around and from five
prime to three prime direction and

bottom Strand are due to this symmetrical
organisation it's believed that a broad.

Similarly fan base pairs
in the center of symmetry.

Must melt to allow
the intermolecular interactions.

So the sea can pair was G.
He will pair was a key and

that process is called extrusion
of cruciform normally.

It doesn't happen.

You need to have certain level
of negative supercommittee and

obviously the composition of nucleotides
is the center of symmetry matters.

So more is due to more easy
this structure will extrude.

So if it happens in one strand
it will lead to formation of

can if it happens in both France and.

Will lead to formation of
so-called cruciform structure.

We decided to look at.

One in the rounds because I pointed
composed of are you repeats.

Are you there are three
hundred base pairs long.

There are a lot of them
in the human genome.

So we have about one million corpus
of the so-called junk D.N.A.

in the human genome which comprises
one percent of the total human D.N.A..

There often are you from clusters
where they're closely spaced so

this is the diagram of human gene and
triangles here.

Arad ones they.

Depict elements in some cases they can
be relative to each other can be in

the rock or in inverted and the like
a certain distance from each other.

Also they are very often the diverge
they are not completely homologous.

So here in lies two G. and which is
next to can one our counter selection

will come to selectable marker
were inserted are you elements.

So created by a very short space or
twelve.

Earth and they differ in the level
of they can be completely homologous

a little bit diverse means that these two
sequences there differ from each other.

And as a control we use absolute the same
repeats indirectly interaction to this if

they cannot adopt any structures and then
how frequently this arm is going to last.

And if you look at look at strains
analyzed trains containing the rattle

Europe is that can not
adopt any structures.

This is how it looks like hope
you can see that this essentially

a place in from each media
to select if media and very.

We can see very few white color in use and

this white color is indicated
in Taishan in can one region.

Happen but
we don't see any red corners here.

Means that in this train.

There are more than is it happens.

This is very very low frequency.

And what if we insert and

work at repeats hundred percent
homologous there you can see drastically.

It's twenty five thousand fold and
look at this microbiology level look

at the score and he is just
completely covered wide popular.

Which indicates that indeed.

The cells they keep losing arm
was incredibly high frequency.

What's amazing about it when I look at
that we're not teaching cells with any.

They need them in the engines.

So we're not teaching them in
the radiation we're not touching that was

any and the nuclear is what we did we just
flipped one repeat relative to each other

and then you have chromosome that
becomes extremely extremely unstable.

This is absolutely in the plan and

it doesn't matter what repeats
you're going to create drama.

You can create confusion of repeats or.

Play it with other gene you're a city or
a top or football in the round effect.

Is the same and
also if we decrease level of

home ology our breaks are still happening
this is happening not as frequently

the case of hundred percent from August
repeats what they see happening and

it happening with quite a high rate.

How this broken chromosomes look like.

If you look at the arrangement of the C.G.

simplistic way of looking if this
chromosome five hundred kilometers long.

If this agent is going to be lost and

its distance between kilometers to
the brake is satisfied you see can be

it's going to become For
this you could be shorter right.

It's a very simple assumption
that he ran it on the geo and

how large the Eagles will.

With the probe.

This is what chromosome five
hundred eighty five could be long

from laughter in the right.

But majority of this chromosomes they run.

Was delayed which means that they not
only lost genetic information but

also again genetic from age.

Except for the school.

How does it happen.

To look at this chromosomes
the structure we use so

called Comparative genome
abuse ational Michael race.

That's a very powerful technique
where you can isolate D.N.A.

from wild strains and
from our strains that last storm

label sure this is the in a label with
different colors mix it one to one ratio.

And expose it to.

When a mike or a swarm of all these
genes aligned here a niche sport.

That's how they're going
to look like they aren't so

each lines here represent promise Sam and
each bar on

top on the boat from you depict
frame this is chromosome

five we're looking east they have sixteen
chromosomes and mitochondria right here.

So the yellow color means there
is no change of copper number.

Blue color it means the region is gone.

Red call it means it's only
present in multiple copies and

this is one of the eyes as you can
see that indeed as we expected.

This thirty could be region is gone.

It turned blue at the same
time right next to it.

There's duplications and there's one
more duplications somewhere else.

On another chromosome.

So if you look at what is common
between these long chromosomes.

If you look at chromosome five.

They're almost identical.

So indeed as we expected based on
our genetic microbiology date or

their become resistant can
they lost their union.

It means that the arm losses and

didn't happen and indeed it happened
at the same time right next to it.

There's a duplications.

If you look at this you can use that
as a signature for this breaks.

So you for invertebrates break.

That's going to look like we're now
a little bit more about this breaks.

We know that.

They.

Way breaks are happening at of inverted P.

is happening very unusual way round
that there's nuclear waste and

we still don't know what is
the advantage of this new place.

It doesn't depend on many and
zines that and.

No in the world that our.

Nucleus of top is in races
the potentially can target the structure.

Nevertheless not of this known players

they cut the structure but
there's someone else.

There's some protein in it
make symmetrical cut and

after the cut is done it's also Or
maybe in conjunction with

maybe it has its own property it
converses broken fragments into hairpins

So it's not the break with the broken and
they're covering the closed.

Completely protected.

But we know that there are not places
that should open this breaks but

if they don't open.

Or sometimes there are too many breaks and
cannot use all of them.

This here they become templates for
the miracle occasion.

If one of the ring or
molecules permanently closed so

you see that both them in opposite
around it can be become a template for

the in Europe location saved in Europe
occasion happens it will not open up

the structure is going to lead to
formation of so-called inverted dimer.

That's that's that's them.

That's the year.

The Freemantle is how it's happening so
inverted is the form cruciform.

It's cleaved semantically when you
form European breaks then they can be

replicated to form on a chromosomal
since this is kilometers.

This is centered near.

This break is going to lead to formation
of a centric fragment centromere fragment.

Since we're following the last
of US centric fragment.

We're going to focus on this.

Central American painting
fragment if this is going.

Escape attack from this nucleus is
limited to formation of dice and

thick and I centric is going to break and
break is going to happen.

Not right in the middle.

It can because it's mechanical break.

This is break form by forces that.

Centromeres to different.

Holes.

It's going to happen randomly and finally.

It's going to be here and

it's going to be healed by edition
of your mirror right here.

Where it can heal if it invades
another chromosome chromosomes usually

we find there's a breakpoint here.

The development Steve
why animals though so

there are also owns that exist in cells.

There are scattered around the genome.

So the can provide a local
level of invasion can happen.

What is interesting about this process we

started from inverted repeat which
is three hundred base pairs alone.

But then we end up with even longer
palindrome which is hundred below.

And if the human

such a geneticist and
if I don't know anything about what I was

happening before they started from and
analyze this chromosome.

Out focus on this break point.

And what I found here.

I'll find you why element.

My conclusion would be that
breaks initiated by elements

is absolutely the wrong conclusion.

Because the break happened.

Hundred could be away and
the culprit the inverted repeat

that initiated this is still sitting
here right in the middle and

this chromosome still unstable they still
can go through the same cycle again.

They can form cruciform it can it can
break it can go and they repeat the cycle.

So the one lesson here is that if you just
analyze breakpoints free range chromosome.

It's not very informative.

So very informative at the same
time this inverted repeats day.

When the break believe
very particular signature.

So the the arm last is
going to be coupled with

duplications inverted
duplications right next to

take a look at the fragment
travelcard the system we can say

can we select who are some some
imagine that it's see me on.

It's like a tip here.

Can the Dr Gene amplification answer is
yes it's also very very fusion process.

So the induction is eleven thousand fold.

That's how the applicants look
like this is normal chromosome.

And if you measure accurately how.

They run we can determine the size and
the Ron and the level of ninety seven Q.B.

although the broken fragment is a fragment
should be forty three could be long.

So knowing the mechanism of this breakage
we suspected that it might be inverted

diameter and the way with that
analysis it's very cool approach.

We used story when you so-called

generic molecular combi
technique coupled with fish.

So this is the achievement of
Georgia Tech mechanical shop they build

this machine four thousand five hundred
dollars X. to begin the last minute.

So how much you want to control
the movement of the arm.

It would diminish so

we can make a mistake this week and if you
have twenty five thousand dollars going to

very very accurate process
how we can go an easy way.

That can be not there some wobbling
here but still it's OK for our purposes.

So what we do we can embark in this
technical was not invented by us.

It's not to.

Our own Bill small slap at POS.

Institute.

So if you can bet your logs.

So you don't share the uni and
then accurately.

Remove fogger rows with our grace and
put it in low ph solution.

And this is cover sleep.

It's here.

And your D.N.A.
is in the solution was low Ph.

Apparently.

Low ph solution.

The only answer for a little bit and

the exposed phosphate groups
that are negatively charged.

On a coward sleep of it's
power to the mind of groups

that are positively charged.

And then we discover
sleep in the solution and

very very slowly pulled out from
the solution what happens that.

The end of chromosomes don't
work who is going to be attached

to the surface of our sleep and then by.

Her surface tension the comma some
are going to be straighten up so

we can see here.

Those are nice chromosomes highlighted and

your story is you're on a cow or sleep.

And you can.

It's a very powerful
technique you can stretch.

Two hundred human genomes
on one hour sleep and

then you can fix the in A and
treat it as essential as membrane.

So that's what we did.

The fish probe which is
a willow both was green and

another problem was the rat.

They're next to each other so
assumption is that if the city engineers

duplicate it if it's in the rocky
between patients going to be red get in.

The red green or it's inverted
intention is going to be red room.

Red Green.

Yes.

And that's exactly what we see.

So this inverted diners
this look exactly like

inverted drivers double
minute that you saw in.

You can.

Very often see in human cancer cells so
that is a sentence fragment.

If it's duplicated it leads
to an inverted diamond from

the discount losing your
conclusion here is that.

This breaks are very powerful way of
generating chromosomal abnormalities.

But the way how this is going to be
visualized depends on selection and

obviously the selection process
has happened in the human cells.

So one one intriguing question is So
we know the consequences

of those breaks the most interesting
is what proteins to make this break.

And in reality when
the structure is formed because

if you imagine just DUPLEIX D.N.A.

The From nation of cruciform exist in
a jet equally in favor of the event.

But it happens when the in
a negative super humidity and

this it can come from transcription or
it can come from the mirror placation So

the approach we used is the classical
approach this is worth the genetics is do

we find mutants and we use two
ways of finding mutants one is so

could you know what scream this
is our impact on system biology.

The reaction that is going
on in in our depart.

And also we tried to use standard
generic screen when we tried to find

mutants using classical genetic approaches
to tell you about genome wide screen.

I'm not going to go into the details
of his genetics is going to be

too overwhelming information but the
bottom line is that several companies they

produce new strains and use genomes
completely sequenced so you can.

They modified each strain in
such a way that they added

that a cycle in promoter to
almost every essential gene and

by society in the media we can deduce
expression level of this essential genes.

Another library we use
a called them library and so

apparently if you introduce a marker
to three prime you two are in big

can of mice and can be a year or
so and isn't it.

Disability and the result that the level

of gene expression goes down
from three to five times.

So we took strains from this library
experience from this library and

look strains from library.

That were each non-essential Gene
was just deleted simply did it not.

Our approach was to take our strain

where we have a system to
the truck chromosomal brakes and

combine them with strains that
carry one of this our levels.

Usually this experiments the done with
robotic systems so this is a system from.

A single company single company
it's produced in England and

it's a very neat way it's very efficient.

So you can.

Team each dog here at represents a colony
and so you can have three hundred ninety.

Six corners of the plate therapeutic
system that can go from one media to

another media and very few this era course
it's a hundred thousand dollars so.

We exported another mushroom from China.

So the name is used.

It's a new generation system.

So she is a very if you sometimes
absolutely amazing person and

much cheaper and
much nicer to interact with us.

So that's she did it in classical way.

So it's amazing but she did six
thousand five hundred crosses and

each time she has to go for me.

That means that she is
sitting on the floor now.

Doing a very nice job trying to cut out
disability and she's using her binge

looked like and
imagine that in a couple of markets.

By herself she did all the jobs and
they aren't She has carried out like this.

So you can see more of a cure you cough
and you can find some mutant mutants

we found out she found almost all of them.

They do want to do in their reputation.

All of those genes if you look like here.

Look what what they do they
felt really and they either.

Essentially this is her draw and
drawing and

she depicted all components of
replication for almost all of them or

almost all of them known that involved in
the leading synthesis of leading strand

strand and if you pick up
any of this D.N.A. proteins.

If you give them then
fragility goes down or

if you mutate so

called replication a city
checkpoint products that that

they feel that sound from Go is
going on wrong in Europe occasion.

So what is our conclusion
conclusion is that.

This brain.

Happening during dinner placation
a say that with the in Arabic

something is going on wrong
during the in Arabic Asian and

the structure occurs but
is still we don't know workers.

This is because the current militants
who are in front of the fourth.

That's who are trying to figure out.

But if you quantity.

The fact.

So this is if you put that this
is under threat promote or

use Mutant the increase in
fragility is quite high.

So this is while typing with.

It's already stone to five
thousand fold over the background.

But if you had a mutation in.

Genes that responsible for synthesis
of Living Color going through and

then fragility goes up and
we can visualize

from the General's Wow So and pulled out a
mutants we can see that chromosomes break.

Normally they aren't using two G.

also it's also very cool technique you can
look at the accumulation of intermediates

during dinner pick ation you can see
that pulled out of mutants they.

This region becomes very problematic for
the synthesis.

The standard approach is
just to give you one minute

of this is to create some damage and

agents that Purify mutant complication
library complement phenotype and

since we can have nice if you doubt we
can go from something like this was

a relatively low level of no fragility or

to hyper fragility we can use this
approach and that was done by.

This totally media and
Natalee with the help of God who is

supposed to destroy all
obstacles in their work and

they were successful they found
mutations in our view to Gene.

And what was.

Amazing about this mutants that some
mutants that they have increased or

decreased so
you can see you break your thousand.

This is a very key player in
the fragility process but

those are a bit too big to use
forms complexes are to be won.

It's part of the promise in the modern
complex it's all they can work on their

own they can be aligned
nicely with one of the Here

it is that the moves will be the junction
and so from structure in bacteria.

In some sense it looks like a little
junction and if you do alignment you can

align and Terminal C. Turmel part
of the proteins with bacterial or

told home all and only you could.

Proteins they have so-called If you
insert so that from the structure.

This is the.

Brown molecule here.

So if you use them we really want
to be to the form either home or

the slice rings and we knew that this

somehow can be loaded through
the cruciform structure and

still in the progress of trying
to understand how it's happening.

And this is of course is trying
to purify the Sprott in some

see what happens with all this mutants and
while invisible.

And we still don't know the simplest
idea that this is still a case that

recognizes the structures and
is trying to remove it from.

From the template.

Or how working more though we think that
there are two pathways one persuade is

going to replication is fine and probably.

Cruciform occurs in front of
replication fork because there's some

local level of negative stupid he.

The city that can drive
the formation of the structure.

It's quite possible that comet
in the modern complex that.

Lately it was found that it travels
along with a replication fork and

might remove his stones.

It can promote the formation
of the structure

at the same time there's
a killer is the should kill this.

Second destruction.

A lot of replication to move forward.

One replication focus the fact that
all the spall the outer Pole all for

Paul Absalom mutants then.

The nation between Legion and
I think that what happens here that

formed on the log in there and then you
have templates and this templates which.

Eventually drives the formation
of course from structure and

most probably the skill is.

Who will work on this
whose forms as well or

not should show a new energy here
on the left on the right but

we have some genetics to prove
it's it's the case one of the.

Challenging things is to try to
detach from nation of course from

structures in the world and nobody's
done it before and the move to working

with Philips intangible has
a very nice probes have

some ideas how to do that and it looks
very sinister at least at this moment.

OK.

What I told you today.

I told you that structures
are very very important

to a new source of chromosome breaks and
obviously this palindromes we can if

you extrapolate the human genome we
can inherit them from parents and

you don't need even to leave extreme
conditions like in Chernobyl area or

during Hiroshima Nagasaki attack but your
chromosomes are going to be predisposed

for breakage if you inherit the second
smarties depending on applied selection.

You can really drive the formation of
your range will see can be either.

Again a lot of a lot of genetic
information the brakes the leaf

specific signature.

So you can even analyze
the results including that you can

use that what is in use in the breaks.

And as I told you what is important for

me is the break points can
be not very informative.

Last couple of new developments.

So our you know why it's in
our pinpoint approach where

genetic backgrounds which we hope we're
going to tell us about the mechanism of

this fragility Textor
political genomes maybe

those are going to be individuals
with the facts that are prone for

this type of rearrangements diseases
this stuff I'm not going to talk.

I don't have time so
I'll just last slide this acknowledgements

Obviously I'll mention some people
who been working on this project and

all of them they did very good job and.

Very tremendous efforts
to make it happen and

we have also new members of the lab
Sabella you would include who joined our

group also like to think
our corporators and support

from funding agencies National Science
Foundation and I thank you very much.

Thank you for your questions.

So really you're going for this nice.

All of the next question
is your next question.

Are also differences.

That's a very interesting question.

So.

If you notice I showed one line.

It's known that translocation happens in
human genome between two palindromes and

this part of Don's the second
on different chromosomes

bartered that it happens only in males.

And it happens with age.

It happens more frequently and
the frequency of this breaks and

formation of Translocation
is one event to ten thousand

cells the analyze this quite a high high
frequent around and definitely there's

definitely a gender and
definitely part of page.

Creations are that it's all
about comets in the morning.

Maybe in their mothers or
the quality in years.

Come back to differently then or
more strongly compacted than all sides and

that compact is ation might influence
the formation of structures and

maybe might influence the breakage.

Well.

We know that already one or
reviewed two are involved.

We know that criminal
remodeling is a player there

do play a role in human
cells this condition.

That's a big question and
definitely like to dress.

As.

Much as.

A good question.

Right.
So is it possible you to them.

Evolution or the US.

If you analyze literature this
know that breaks not happening.

Ground.

It came from evolution the star
the sole parent Lee if you know lies.

You know so
you can say you can see somebody

genes that evolve faster or slower and

more nickel view was that all is
coming from genetic mutations.

So apparently it's not the case anymore.

Currently breakage and rearranged it's
a very powerful way to drive evolution and

the simulation is that the selection and

what is also interesting that apparently
the rate of breakage is different

from regions that contain
actively transcribed genes and

those that are more silent and those of
the most active genes the break faster.

So in this way from evolutionary or

from work to point a few you want
decisions to evolve faster as well.

And other ways.

How dense are you so.

This formation of double minute.

Is quite a big problem cancer therapy.

It's one of the ways how cancer cells
they can acquire resistance to drugs.

Imagine that some gene that has potential

to block the activity of
the chemicals would be amplified.

Then you're going to shield
yourself from this activity so

if you under five players
that generate this break.

That is a potential target for
the cam for the therapy.

So if you combine year.

Came up in a political heat with agent
that was going to block the nucleus

of the going to block
factors the promote breakage.

Maybe your chances of relapse
going to the US still.

Just to expand the.

Father for human status.

Yes.

You know you have your so

normal technology with global sequence in
that it's getting more and more valuable.

So that was what you mentioned during
my talk with the scope a number of very

becomes very popular much here in
the human genetic study is that

even what you cannot find
using the spectral type and

still would differ from each other and
were different from each other.

Not only at the basis of point
mutations and small changes.

They also were different from each other
at the level of how many copies of

particular genes and how many copies
it means genomic rearrangement so that

not drastic general careers that are going
to completely change the architecture of

chromosome but rather change the local
context local level of expression.

That's definitely a factor and breaks and
sequences that promote this breaks.

I think those are quite important
players in this process.

Like yours.

And thank you thanks.