Generation of a TLE3 heterozygous knockout human embryonic stem cell line using CRISPR-Cas9

Anne M Bara; Angelica Messana; Amanda Herring; Dane Z Hazelbaker; Kevin Eggan; Lindy E Barrett

doi:10.1016/j.scr.2016.09.008

Generation of a TLE3 heterozygous knockout human embryonic stem cell line using CRISPR-Cas9

Stem Cell Res. 2016 Sep;17(2):441-443. doi: 10.1016/j.scr.2016.09.008. Epub 2016 Sep 15.

Authors

Anne M Bara¹, Angelica Messana¹, Amanda Herring¹, Dane Z Hazelbaker¹, Kevin Eggan², Lindy E Barrett³

Affiliations

¹ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
² Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA.
³ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA. Electronic address: lbarrett@broadinstitute.org.

PMID: 27879221
DOI: 10.1016/j.scr.2016.09.008

Abstract

Here, we generated a monoallelic mutation in the TLE3 (Transducin Like Enhancer of Split 3) gene using CRISPR-Cas9 editing in the human embryonic stem cell (hESC) line WA01. The heterozygous knockout cell line, TLE3-447-D08-A01, displays partial loss of TLE3 protein expression while maintaining pluripotency, differentiation potential and genomic integrity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
Blotting, Western
CRISPR-Cas Systems / genetics*
Cell Line
Co-Repressor Proteins / chemistry
Co-Repressor Proteins / genetics*
Co-Repressor Proteins / metabolism
Embryoid Bodies / metabolism
Embryoid Bodies / pathology
Heterozygote
Human Embryonic Stem Cells
Humans
Karyotype
Real-Time Polymerase Chain Reaction
Transcription Factors / genetics
Transcription Factors / metabolism

Substances

Co-Repressor Proteins
TLE3 protein, human
Transcription Factors