A method for genome editing in human pluripotent stem cells

Cory Smith, Zhaohui Ye, Linzhao Cheng

Research output: Contribution to journalArticlepeer-review

Abstract

Human pluripotent stem cells (PSCs) hold great potential for regenerative medicine and currently are being used as a research tool for basic discovery and disease modeling. To evaluate the role of a single genetic variant, a system of genome editing is needed to precisely mutate any desired DNA sequence in isolation and measure its effect on phenotype when compared to the isogenic parental PSC from which it was derived. This protocol describes the general targeting schemes used by researchers to edit PSCs to knock out, knock-in, or precisely alter a single nucleotide, covering conditions for electroporation, clonal isolation, and screening of edited PSCs for the targeted mutation. These recent advances simplify the procedure for genome editing, allowing individual researchers to induce nearly any desired mutation to further study its function or to reverse a disease-causing variant for future applications in regenerative medicine.

Original languageEnglish (US)
Pages (from-to)385-390
Number of pages6
JournalCold Spring Harbor Protocols
Volume2016
Issue number4
DOIs
StatePublished - Apr 1 2016

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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