TY - JOUR
T1 - A method for genome editing in human pluripotent stem cells
AU - Smith, Cory
AU - Ye, Zhaohui
AU - Cheng, Linzhao
N1 - Publisher Copyright:
© 2016 Cold Spring Harbor Laboratory Press.
PY - 2016/4
Y1 - 2016/4
N2 - 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.
AB - 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.
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U2 - 10.1101/pdb.prot090217
DO - 10.1101/pdb.prot090217
M3 - Article
C2 - 27037073
AN - SCOPUS:84962419845
SN - 1940-3402
VL - 2016
SP - 385
EP - 390
JO - Cold Spring Harbor Protocols
JF - Cold Spring Harbor Protocols
IS - 4
ER -