scholarly journals Single-Step Generation of Conditional Knockout Mouse Embryonic Stem Cells

Cell Reports ◽  
2015 ◽  
Vol 12 (4) ◽  
pp. 709-716 ◽  
Author(s):  
Matyas Flemr ◽  
Marc Bühler
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Knockout Mouse ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Single Step ◽  
Conditional Knockout ◽  
Conditional Knockout Mouse ◽  
Step Generation

scholarly journals Selection-Independent Generation of Gene Knockout Mouse Embryonic Stem Cells Using Zinc-Finger Nucleases

PLoS ONE ◽  
2011 ◽  
Vol 6 (12) ◽  
pp. e28911 ◽  
Author(s):  
Anna Osiak ◽  
Frank Radecke ◽  
Eva Guhl ◽  
Sarah Radecke ◽  
Nadine Dannemann ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Knockout Mouse ◽  
Gene Knockout ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Zinc Finger Nucleases ◽  
Gene Knockout Mouse

Rapid Single-Step Separation of Pluripotent Mouse Embryonic Stem Cells from Mouse Feeder Fibroblasts

2008 ◽  
Vol 17 (2) ◽  
pp. 383-388 ◽  
Author(s):  
Zhixin Li ◽  
Matthew R. Barron ◽  
John Lough ◽  
Ming Zhao
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Single Step

Cre-mediated deletion of SMARCA5 disrupts pluripotency in mouse embryonic stem cells

2020 ◽  
Author(s):  
David P. Cook ◽  
Barbara C. Vanderhyden
Keyword(s):  
Stem Cells ◽  
Transcription Factors ◽  
Embryonic Stem Cells ◽  
Binding Sites ◽  
Morphological Changes ◽  
Embryonic Stem ◽  
Conditional Knockout ◽  
Mouse Escs ◽  
Conditional Knockout Mouse ◽  
Genome Wide

AbstractIn embryonic stem cells (ESCs), the SWI/SNF, CHD, and INO80 families of ATP-dependent chromatin remodellers have been implicated in maintaining pluripotency-associated gene expression. At the time of this study, the importance of ISWI family remodellers had yet to be defined, and we had sought to assess their involvement. During this time, Barisic et al. (Nature, 2019) elegantly demonstrated that the ISWI homologue SNF2H (Smarca5) is important for nucleosomal periodicity, the binding of select transcription factors, and proper differentiation of mouse ESCs. While we do not dispute their findings to any extent, our experiments have led to slightly different conclusions, and we have chosen to use this platform to share our results.Here, we explore the importance of SNF2H by deriving a conditional knockout mouse ESC line and observing the consequences of SNF2H depletion on the pluripotent state. Cre-mediated deletion of Snf2h disrupts hallmark characteristics of pluripotency, resulting in distinct morphological changes; reduced expression of the master transcription factors Oct4, Sox2, and Nanog; and reduced alkaline phosphatase activity. To understand the mechanisms of SNF2H-mediated regulation, we mapped SNF2H-bound nucleosomes genome-wide. SNF2H is broadly distributed across the genome but is preferentially enriched at active regulatory regions and transcription factor binding sites.

scholarly journals Enhanced Genomic Instability and Defective Postreplication Repair in RAD18 Knockout Mouse Embryonic Stem Cells

2003 ◽  
Vol 23 (2) ◽  
pp. 474-481 ◽  
Author(s):  
Satoshi Tateishi ◽  
Hitoshi Niwa ◽  
Jun-Ichi Miyazaki ◽  
Shiho Fujimoto ◽  
Hirokazu Inoue ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Gene Targeting ◽  
Knockout Mouse ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells ◽  
Human Cells ◽  
Wild Type ◽  
Normal Cells ◽  
Postreplication Repair

ABSTRACT In lower eukaryotes, Rad18 plays a crucial role in postreplication repair. Previously, we isolated a human homologue of RAD18 (hRAD18) and showed that human cells overexpressing hRad18 protein with a mutation in the RING finger motif are defective in postreplication repair. Here, we report the construction of RAD18-knockout mouse embryonic stem cells by gene targeting. These cells had almost the same growth rate as wild-type cells and manifested phenotypes similar to those of human cells expressing mutant Rad18 protein: hypersensitivity to multiple DNA damaging agents and a defect in postreplication repair. Mutation was not induced in the knockout cells with any higher frequencies than in wild-type cells, as shown by ouabain resistance. In the knockout cells, spontaneous sister chromatid exchange (SCE) occurred with twice the frequency observed in normal cells. After mild DNA damage, SCE was threefold higher in the knockout cells, while no increase was observed in normal cells. Stable transformation efficiencies were ∼20-fold higher in knockout cells, and gene targeting occurred with ∼40-fold-higher frequency than in wild-type cells at the Oct3/4 locus. These results indicate that dysfunction of Rad18 greatly increases both the frequency of homologous as well as illegitimate recombination, and that RAD18 contributes to maintenance of genomic stability through postreplication repair.

Sign in / Sign up

Export Citation Format

Share Document