Gene Replacement of the p53 Gene with the lacZ Gene in Mouse Embryonic Stem-Cells and Mice by Using Two Steps of Homologous Recombination

1994 ◽  
Vol 202 (2) ◽  
pp. 830-837 ◽  
Author(s):  
Y. Gondo ◽  
K. Nakamura ◽  
K. Nakao ◽  
T. Sasaoka ◽  
K. Ito ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Homologous Recombination ◽  
Embryonic Stem ◽  
P53 Gene ◽  
Gene Replacement ◽  
Mouse Embryonic Stem Cells ◽  
Lacz Gene

scholarly journals The length of homology required for gene targeting in embryonic stem cells

1991 ◽  
Vol 11 (11) ◽  
pp. 5586-5591 ◽  
Author(s):  
P Hasty ◽  
J Rivera-Pérez ◽  
A Bradley
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Homologous Recombination ◽  
Gene Targeting ◽  
Es Cells ◽  
Embryonic Stem ◽  
Critical Length ◽  
Gene Replacement ◽  
Site Specific ◽  
The Relationship

Homologous recombination has been used to introduce site-specific mutations into murine embryonic stem (ES) cells with both insertion and replacement vectors. In this study, we compared the frequency of gene targeting with various lengths of homology and found a dramatic increase in targeting with an increase in homology from 1.3 to 6.8 kb. We examined in detail the relationship between the length of homology and the gene-targeting frequency for replacement vectors and found that a critical length of homology is needed for targeting. Adding greater lengths of homology to this critical length has less of an effect on the targeting frequency. We also analyzed the lengths of homology necessary on both arms of the vector for gene replacement events and found that 472 bp of homology is used as efficiently as 1.2 kb in the formation and resolution of crossover junctions.

scholarly journals RTEL1 contributes to DNA replication and repair and telomere maintenance

2012 ◽  
Vol 23 (14) ◽  
pp. 2782-2792 ◽  
Author(s):  
Evert-Jan Uringa ◽  
Kathleen Lisaingo ◽  
Hilda A. Pickett ◽  
Julie Brind'Amour ◽  
Jan-Hendrik Rohde ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dna Repair ◽  
Dna Replication ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Gene Replacement ◽  
Mouse Embryonic Stem Cells ◽  
Telomere Maintenance ◽  
Dominant Factor ◽  
Deficient Mouse

Telomere maintenance and DNA repair are important processes that protect the genome against instability. mRtel1, an essential helicase, is a dominant factor setting telomere length in mice. In addition, mRtel1 is involved in DNA double-strand break repair. The role of mRtel1 in telomere maintenance and genome stability is poorly understood. Therefore we used mRtel1-deficient mouse embryonic stem cells to examine the function of mRtel1 in replication, DNA repair, recombination, and telomere maintenance. mRtel1-deficient mouse embryonic stem cells showed sensitivity to a range of DNA-damaging agents, highlighting its role in replication and genome maintenance. Deletion of mRtel1 increased the frequency of sister chromatid exchange events and suppressed gene replacement, demonstrating the involvement of the protein in homologous recombination. mRtel1 localized transiently at telomeres and is needed for efficient telomere replication. Of interest, in the absence of mRtel1, telomeres in embryonic stem cells appeared relatively stable in length, suggesting that mRtel1 is required to allow extension by telomerase. We propose that mRtel1 is a key protein for DNA replication, recombination, and repair and efficient elongation of telomeres by telomerase.

scholarly journals Production of p53 gene knockout rats by homologous recombination in embryonic stem cells

Nature ◽  
2010 ◽  
Vol 467 (7312) ◽  
pp. 211-213 ◽  
Author(s):  
Chang Tong ◽  
Ping Li ◽  
Nancy L. Wu ◽  
Youzhen Yan ◽  
Qi-Long Ying
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Homologous Recombination ◽  
Gene Knockout ◽  
Embryonic Stem ◽  
P53 Gene
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