Efficient scarless genome editing in human pluripotent stem cells

Kazuya Ikeda; Nobuko Uchida; Toshinobu Nishimura; Joseph White; Renata M Martin; Hiromitsu Nakauchi; Vittorio Sebastiano; Kenneth I Weinberg; Matthew H Porteus

doi:10.1038/s41592-018-0212-y

Efficient scarless genome editing in human pluripotent stem cells

Nat Methods. 2018 Dec;15(12):1045-1047. doi: 10.1038/s41592-018-0212-y. Epub 2018 Nov 30.

Authors

Kazuya Ikeda¹, Nobuko Uchida², Toshinobu Nishimura³, Joseph White⁴, Renata M Martin¹, Hiromitsu Nakauchi³, Vittorio Sebastiano⁴, Kenneth I Weinberg¹, Matthew H Porteus⁵

Affiliations

¹ Department of Pediatrics, Stanford University, Stanford, CA, USA.
² ReGen Med Division, BOCO Silicon Valley, Palo Alto, CA, USA.
³ Department of Genetics, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA.
⁴ Department of Obstetrics and Gynecology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, Stanford, CA, USA.
⁵ Department of Pediatrics, Stanford University, Stanford, CA, USA. mporteus@stanford.edu.

PMID: 30504872
DOI: 10.1038/s41592-018-0212-y

Abstract

Scarless genome editing in human pluripotent stem cells (hPSCs) represents a goal for both precise research applications and clinical translation of hPSC-derived therapies. Here we established a versatile and efficient method that combines CRISPR-Cas9-mediated homologous recombination with positive-negative selection of edited clones to generate scarless genetic changes in hPSCs.

Publication types

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

MeSH terms

CRISPR-Cas Systems*
Embryonic Stem Cells / cytology
Embryonic Stem Cells / metabolism*
Gene Editing*
Gene Expression Regulation
Genome, Human*
Homologous Recombination*
Humans
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism*
RNA, Small Interfering / genetics*

Substances

RNA, Small Interfering