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Human targeted deletions and biologi...

Ahmad, Alya.

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Human targeted deletions and biological roles of genes involved in repair of alkylation damage.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Human targeted deletions and biological roles of genes involved in repair of alkylation damage./
Author:	Ahmad, Alya.
Description:	76 p.
Notes:	Source: Masters Abstracts International, Volume: 54-05.
Contained By:	Masters Abstracts International54-05(E).
Subject:	Cellular biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=1591663
ISBN:	9781321835090

Human targeted deletions and biological roles of genes involved in repair of alkylation damage.
Ahmad, Alya.

Human targeted deletions and biological roles of genes involved in repair of alkylation damage. - 76 p.

Source: Masters Abstracts International, Volume: 54-05.

Thesis (M.S.)--Boston University, 2015.

DNA repair is not a single mechanism found within cells. There exists numerous different DNA repair mechanisms that function within every type of cell. The majority of these mechanisms risk accumulating mutations. However, there are a few repair mechanisms that are known to be error-free and one of these is direct reversal repair. This study focused on two proteins highly involved in direct reversal DNA repair--ALKBH2 and ALKBH3. Previous studies have shown that in mice, these two proteins play a significant role in preventing and repairing DNA damage due to methylation as well as decreasing the frequency of mutagenic alkyl adducts. The goal of this study was to characterize the roles of the direct reversal repair proteins in human cells. We expected to see a similar phenotype to that of the Alkbh2 and Alkbh3-deficient mice. Telomerase immortalized human skin fibroblasts were targeted for the ALKBH2 and ALKBH3 alleles using a RNA-guided CRISPR-Cas9 construct that was designed to induce double stranded DNA breaks within the exons and disrupt the open reading frame, eliminating protein activity. Isolated clones were analyzed using fragment analysis and DNA sequencing to characterize any alterations in the open reading frame of the genes. Through sequencing analysis, results showed that one clone was successfully targeted for one of the ALKBH3 alleles with a single nucleotide insertion in its sequence, causing a disruption of the open reading frame. Though the ultimate goal of the experiment was not attained, we concluded that HTERTG fibroblasts can be expanded to serve as a model in which to construct targeted human cell lines that have near normal karyotypes.

ISBN: 9781321835090Subjects--Topical Terms:

3172791
Cellular biology.

Human targeted deletions and biological roles of genes involved in repair of alkylation damage.
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245 1 0 $a Human targeted deletions and biological roles of genes involved in repair of alkylation damage.
300 $a 76 p.
500 $a Source: Masters Abstracts International, Volume: 54-05.
500 $a Adviser: R. Jarrett Rushmore III.
502 $a Thesis (M.S.)--Boston University, 2015.
520 $a DNA repair is not a single mechanism found within cells. There exists numerous different DNA repair mechanisms that function within every type of cell. The majority of these mechanisms risk accumulating mutations. However, there are a few repair mechanisms that are known to be error-free and one of these is direct reversal repair. This study focused on two proteins highly involved in direct reversal DNA repair--ALKBH2 and ALKBH3. Previous studies have shown that in mice, these two proteins play a significant role in preventing and repairing DNA damage due to methylation as well as decreasing the frequency of mutagenic alkyl adducts. The goal of this study was to characterize the roles of the direct reversal repair proteins in human cells. We expected to see a similar phenotype to that of the Alkbh2 and Alkbh3-deficient mice. Telomerase immortalized human skin fibroblasts were targeted for the ALKBH2 and ALKBH3 alleles using a RNA-guided CRISPR-Cas9 construct that was designed to induce double stranded DNA breaks within the exons and disrupt the open reading frame, eliminating protein activity. Isolated clones were analyzed using fragment analysis and DNA sequencing to characterize any alterations in the open reading frame of the genes. Through sequencing analysis, results showed that one clone was successfully targeted for one of the ALKBH3 alleles with a single nucleotide insertion in its sequence, causing a disruption of the open reading frame. Though the ultimate goal of the experiment was not attained, we concluded that HTERTG fibroblasts can be expanded to serve as a model in which to construct targeted human cell lines that have near normal karyotypes.
590 $a School code: 0017.
650 4 $a Cellular biology. $3 3172791
650 4 $a Oncology. $3 751006
650 4 $a Biology. $3 522710
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710 2 $a Boston University. $b Medical Sciences. $3 2104756
773 0 $t Masters Abstracts International $g 54-05(E).
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791 $a M.S.
792 $a 2015
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=1591663