rapid generation of human genetic loss-of-function ipsc lines by simultaneous reprogramming and gene editing

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ID: 205429
2017
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Abstract
Specifically ablating genes in human induced pluripotent stem cells (iPSCs) allows for studies of gene function as well as disease mechanisms in disorders caused by loss-of-function (LOF) mutations. While techniques exist for engineering such lines, we have developed and rigorously validated a method of simultaneous iPSC reprogramming while generating CRISPR/Cas9-dependent insertions/deletions (indels). This approach allows for the efficient and rapid formation of genetic LOF human disease cell models with isogenic controls. The rate of mutagenized lines was strikingly consistent across experiments targeting four different human epileptic encephalopathy genes and a metabolic enzyme-encoding gene, and was more efficient and consistent than using CRISPR gene editing of established iPSC lines. The ability of our streamlined method to reproducibly generate heterozygous and homozygous LOF iPSC lines with passage-matched isogenic controls in a single step provides for the rapid development of LOF disease models with ideal control lines, even in the absence of patient tissue.
Reference Key
tidball2017stemrapid Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors ;Andrew M. Tidball;Louis T. Dang;Trevor W. Glenn;Emma G. Kilbane;Daniel J. Klarr;Joshua L. Margolis;Michael D. Uhler;Jack M. Parent
Journal nature reviews gastroenterology & hepatology
Year 2017
DOI
10.1016/j.stemcr.2017.07.003
URL
http://www.sciencedirect.com/science/article/pii/S2213671117303132
Keywords
induced pluripotent stem cells reprogramming crispr/cas9 gene editing

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