Generation of a TLE1 homozygous knockout human embryonic stem cell line using CRISPR-Cas9

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

doi:10.1016/j.scr.2016.09.009

Generation of a TLE1 homozygous knockout human embryonic stem cell line using CRISPR-Cas9

Stem Cell Res. 2016 Sep;17(2):430-432. doi: 10.1016/j.scr.2016.09.009. Epub 2016 Sep 16.

Authors

Amanda Herring¹, Angelica Messana¹, Anne M Bara¹, 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: 27879218
DOI: 10.1016/j.scr.2016.09.009

Abstract

Here, we generated a biallelic mutation in the TLE1 (Transducin Like Enhancer of Split 1) gene using CRISPR-Cas9 editing in the human embryonic stem cell (hESC) line WA01. The homozygous knockout cell line, TLE1-464-G04, displays loss of TLE1 protein expression while maintaining pluripotency, differentiation potential and genomic integrity.

MeSH terms

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

Substances

Co-Repressor Proteins
Repressor Proteins
TLE1 protein, human
Transcription Factors