Title:
Establishing a Working Protocol for Plasmid Cloning and shRNA Design in Endogenous Brachionus manjavacas Gene TRP7

dc.contributor.advisor Snell, Terry
dc.contributor.author Krishnappan, Sharadha
dc.contributor.committeeMember Hammer, Brian
dc.contributor.department Biology
dc.date.accessioned 2016-07-18T17:05:37Z
dc.date.available 2016-07-18T17:05:37Z
dc.date.created 2016-05
dc.date.issued 2016-07-18
dc.date.submitted May 2016
dc.date.updated 2016-07-18T17:05:37Z
dc.description.abstract Current transfection protocol in rotifers only allows for temporary transfection within rotifers and does not allow for the continuous knockdown of endogenous genes, thereby inhibiting the possibility of observing long-term biological effects in response to specific perpetual gene knockdowns. This study aims to address this particular issue by establishing a working protocol for plasmid cloning and shRNA design within an endogenous gene of B. manjavacas with known biological effects, allowing for the exploration into the optimization of a transfection protocol and demonstration of RNAi knockdown of the known gene within the rotifers as subsequent studies. Manipulation of gene expression in rotifers could occur through plasmid vector insertions, which induce silencing of a gene’s expression with short hairpin RNA (shRNA), via RNAi. This would effectively stimulate gene knockdown, allowing for the observation of biological effects such as changes in fecundity and lifespan. With the establishment of a working protocol for plasmid cloning and shRNA design, as a result of this study, the optimization of a transfection protocol for rotifers is explored. With increased efficiency in the transfection of rotifers, populations of rotifers expressing the plasmid can be amassed, allowing for experimental design that examine the varying aging mechanisms and effects that are stimulated due to permanent changes in target gene expression through RNAi. This, in turn, could give rise to the identification of evolutionarily conserved genes that regulate organismal aging, which could lead to further implications in the field of pharmacological intervention in mammalian aging as well as in the field of biogerontology overall.
dc.description.degree Undergraduate
dc.format.mimetype application/pdf
dc.identifier.uri http://hdl.handle.net/1853/55402
dc.language.iso en_US
dc.publisher Georgia Institute of Technology
dc.subject Plasmid cloning
dc.subject shRNA Design
dc.subject Rotifer
dc.subject Brachionus manjavacas
dc.subject TRP7
dc.subject Short hairpin RNA
dc.subject Biogerontology
dc.subject Transfection
dc.subject Gene knockdown
dc.subject RNAi knockdown
dc.title Establishing a Working Protocol for Plasmid Cloning and shRNA Design in Endogenous Brachionus manjavacas Gene TRP7
dc.type Text
dc.type.genre Undergraduate Thesis
dspace.entity.type Publication
local.contributor.corporatename College of Sciences
local.contributor.corporatename School of Biological Sciences
local.contributor.corporatename Undergraduate Research Opportunities Program
local.relation.ispartofseries Undergraduate Research Option Theses
relation.isOrgUnitOfPublication 85042be6-2d68-4e07-b384-e1f908fae48a
relation.isOrgUnitOfPublication c8b3bd08-9989-40d3-afe3-e0ad8d5c72b5
relation.isOrgUnitOfPublication 0db885f5-939b-4de1-807b-f2ec73714200
relation.isSeriesOfPublication e1a827bd-cf25-4b83-ba24-70848b7036ac
thesis.degree.level Undergraduate
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