scholarly journals Gene Editing of Human Embryonic Stem Cells via an Engineered Baculoviral Vector Carrying Zinc-finger Nucleases

2011 ◽  
Vol 19 (5) ◽  
pp. 942-950 ◽  
Author(s):  
Yuning Lei ◽  
Chi-Lin Lee ◽  
Kye-Il Joo ◽  
Jonathan Zarzar ◽  
Yarong Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Human Embryonic Stem Cells ◽  
Gene Editing ◽  
Embryonic Stem ◽  
Zinc Finger Nucleases

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

scholarly journals ZNF91 deletion in human embryonic stem cells leads to ectopic activation of SVA retrotransposons and up-regulation of KRAB zinc finger gene clusters

2021 ◽  
Author(s):  
Nina L. Haring ◽  
Elisabeth J. van Bree ◽  
Whitney S. Jordaan ◽  
Judith R.E. Roels ◽  
Gonzalo Congrains Sotomayor ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Gene Clusters ◽  
Zinc Finger Gene ◽  
Ectopic Activation

scholarly journals Functional interrogation of Lynch syndrome‐associated MSH2 missense variants via CRISPR‐Cas9 gene editing in human embryonic stem cells

2019 ◽  
Vol 40 (11) ◽  
pp. 2044-2056 ◽  
Author(s):  
Abhijit Rath ◽  
Akriti Mishra ◽  
Victoria Duque Ferreira ◽  
Chaoran Hu ◽  
Gregory Omerza ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Lynch Syndrome ◽  
Human Embryonic Stem Cells ◽  
Gene Editing ◽  
Embryonic Stem ◽  
Missense Variants

scholarly journals Zinc finger E-box–binding homeobox 1 (ZEB1) is required for neural differentiation of human embryonic stem cells

2018 ◽  
Vol 293 (50) ◽  
pp. 19317-19329 ◽  
Author(s):  
Yuan Jiang ◽  
Long Yan ◽  
Longkuo Xia ◽  
Xiaoyin Lu ◽  
Wenliang Zhu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Human Embryonic Stem Cells ◽  
Neural Differentiation ◽  
Embryonic Stem ◽  
E Box

scholarly journals Gene editing in haploid human embryonic stem cells using CRISPR/Cas9

2017 ◽  
Vol 108 (3) ◽  
pp. e24
Author(s):  
L.Z. Safier ◽  
M.V. Zuccaro ◽  
R. Lobo ◽  
D. Egli
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Gene Editing ◽  
Embryonic Stem

scholarly journals Functional Interrogation of Lynch Syndrome Associated MSH2 Missense Variants Using CRISPR-Cas9 Gene Editing in Human Embryonic Stem Cells

2018 ◽  
Author(s):  
Abhijit Rath ◽  
Akriti Mishra ◽  
Victoria Duque Ferreira ◽  
Chaoran Hu ◽  
James P. Grady ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Lynch Syndrome ◽  
Human Embryonic Stem Cells ◽  
Gene Editing ◽  
Embryonic Stem ◽  
Functional Assay ◽  
Missense Variants ◽  
Mmr Genes ◽  
Artificial Nature

AbstractLynch syndrome (LS) is a hereditary cancer predisposition condition caused by inactivating germline mutations in one of the DNA mismatch repair (MMR) genes. Identifying a deleterious germline mutation by DNA sequencing is important for confirming an LS diagnosis. Frameshift and nonsense mutations significantly alter the protein product and likely impair MMR function. However, the implication of a missense mutation is often difficult to interpret. Referred to as variants of uncertain significance (VUS), their discovery hampers the definitive LS diagnosis. To determine the pathogenic significance of a VUS it is helpful to know its impact on protein function. Functional studies in the test tube and in cellular models have been performed for some VUS, however, these studies have been limited by the artificial nature of the assays. We report here an improved functional assay in which we engineered site-specific MSH2 VUS using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 gene editing in human embryonic stem cells. This approach introduces the variant into the endogenous MSH2 loci, while simultaneously eliminating the wild-type gene. We then characterized the impact of the variants on cellular MMR functions including DNA damage response signaling upon challenge with a DNA alkylating agent and the repair of DNA microsatellites. We classified the MMR functional capability of 8 of 10 VUS under study providing valuable information for determining their likelihood of being bona fide LS mutations. This improved human cell-based assay system for functionally testing MMR gene VUS will facilitate the identification of high risk LS patients.Significance StatementUnderstanding how cancer-associated missense variants in MMR genes affect function helps determine whether they truly contribute to disease. Laboratory assays previously utilized are limited by their artificial nature. To improve this, we introduced variants directly into the endogenous MMR loci in hESCs using CRISPR-Cas9 gene editing. This approach allows us to assess each variant while being expressed by its normal regulatory elements in a cellular environment. Our results will help guide the management of patients world-wide who carry these variants. At the same time, this study provides a technical road map for assessing the functional effects of all LS-associated variants, as well as variants linked to other genetic diseases where a cell-based functional assay is available.

Resistance to zinc finger nuclease (ZFN)-mediated FANCA disruption in human embryonic stem cells (hESC)

2013 ◽  
Vol 41 (8) ◽  
pp. S29
Author(s):  
Kim Vanuytsel ◽  
Laura Ordovas Vidal ◽  
Qing Cai ◽  
Nisha Nair ◽  
Rob Van Rossom ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Zinc Finger ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Zinc Finger Nuclease
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