2015 Dec 8;5(6):1097-1108. doi: 10.1016/j.stemcr.2015.10.010. Epub 2015 Nov 19.

Author information

1
Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA.
2
A-T Clinic, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
3
Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA; Department of Genetics, Rutgers University, Piscataway, NJ 08854, USA; Human Genetics Institute of New Jersey, Piscataway, NJ 08854, USA.
4
Department of Cell Biology and Neuroscience, Rutgers University, Piscataway, NJ 08854, USA; Human Genetics Institute of New Jersey, Piscataway, NJ 08854, USA. Electronic address: rhart@rutgers.edu.

Abstract

A spontaneously reverted iPSC line was identified from an A-T subject with heterozygous ATM truncation mutations. The reverted iPSC line expressed ATM protein and was capable of radiation-induced phosphorylation of CHK2 and H2A.X. Genome-wide SNP analysis confirmed a match to source T cells and also to a distinct, non-reverted iPSC line from the same subject. Rearranged T cell receptor sequences predict that the iPSC culture originated as several independently reprogrammed cells that resolved into a single major clone, suggesting that gene correction likely occurred early in the reprogramming process. Gene expression analysis comparing ATM(-/-) iPSC lines to unrelated ATM(+/-) cells identifies a large number of differences, but comparing only the isogenic pair of A-T iPSC lines reveals that the primary pathway affected by loss of ATM is a diminished expression of p53-related mRNAs. Gene reversion in culture, although likely a rare event, provided a novel, reverted cell line for studying ATM function.

PMID:
 
26677768
 
PMCID:
 
PMC4682125
 
DOI:
 
10.1016/j.stemcr.2015.10.010
[Indexed for MEDLINE] 
Free PMC Article