scholarly journals The structure-specific endonuclease Ercc1-Xpf is required for targeted gene replacement in embryonic stem cells

2001 ◽  
Vol 20 (22) ◽  
pp. 6540-6549 ◽  
Author(s):  
L. J. Niedernhofer
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Gene Replacement ◽  
Targeted Gene Replacement

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 Erratum: Corrigendum: Mitochondrial gene replacement in primate offspring and embryonic stem cells

Nature ◽  
2014 ◽  
Vol 516 (7530) ◽  
pp. 276-276
Author(s):  
Masahito Tachibana ◽  
Michelle Sparman ◽  
Hathaitip Sritanaudomchai ◽  
Hong Ma ◽  
Lisa Clepper ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Mitochondrial Gene ◽  
Embryonic Stem ◽  
Gene Replacement

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 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 I-SceI-Induced Gene Replacement at a Natural Locus in Embryonic Stem Cells

1998 ◽  
Vol 18 (3) ◽  
pp. 1444-1448 ◽  
Author(s):  
Michel Cohen-Tannoudji ◽  
Sylvie Robine ◽  
André Choulika ◽  
Daniel Pinto ◽  
Fatima El Marjou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Targeting ◽  
Mammalian Cells ◽  
High Efficiency ◽  
Shuttle Vector ◽  
Embryonic Stem ◽  
Gene Replacement ◽  
Actin Binding ◽  
Selection Marker

ABSTRACT Gene targeting is a very powerful tool for studying mammalian development and physiology and for creating models of human diseases. In many instances, however, it is desirable to study different modifications of a target gene, but this is limited by the generally low frequency of homologous recombination in mammalian cells. We have developed a novel gene-targeting strategy in mouse embryonic stem cells that is based on the induction of endogenous gap repair processes at a defined location within the genome by induction of a double-strand break (DSB) in the gene to be mutated. This strategy was used to knock in an NH2-ezrin mutant in the villin gene, which encodes an actin-binding protein expressed in the brush border of the intestine and the kidney. To induce the DSB, an I-SceI yeast meganuclease restriction site was first introduced by gene targeting to the villin gene, followed by transient expression of I-SceI. The repair of the ensuing DSB was achieved with high efficiency (6 × 10−6) by a repair shuttle vector sharing only a 2.8-kb region of homology with the villin gene and no negative selection marker. Compared to conventional gene-targeting experiments at the villin locus, this represents a 100-fold stimulation of gene-targeting frequency, notwithstanding a much lower length of homology. This strategy will be very helpful in facilitating the targeted introduction of several types of mutations within a gene of interest.

scholarly journals Mitochondrial gene replacement in primate offspring and embryonic stem cells

Nature ◽  
2009 ◽  
Vol 461 (7262) ◽  
pp. 367-372 ◽  
Author(s):  
Masahito Tachibana ◽  
Michelle Sparman ◽  
Hathaitip Sritanaudomchai ◽  
Hong Ma ◽  
Lisa Clepper ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Mitochondrial Gene ◽  
Embryonic Stem ◽  
Gene Replacement
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