A stress-free strategy to correct point mutations in patient iPS cells.
CRISPR-Cas9
Gene targeting
Isogenic iPS cells
Point mutation
Journal
Stem cell research
ISSN: 1876-7753
Titre abrégé: Stem Cell Res
Pays: England
ID NLM: 101316957
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
received:
22
10
2020
revised:
23
02
2021
accepted:
01
04
2021
pubmed:
16
4
2021
medline:
1
7
2021
entrez:
15
4
2021
Statut:
ppublish
Résumé
When studying patient specific induced pluripotent stem cells (iPS cells) as a disease model, the ideal control is an isogenic line that has corrected the point mutation, instead of iPS cells from siblings or other healthy subjects. However, repairing a point mutation in iPS cells even with the newly developed CRISPR-Cas9 technique remains difficult and time-consuming. Here we report a strategy that makes the Cas9 "knock-in" methodology both hassle-free and error-free. Instead of selecting a Cas9 recognition site close to the point mutation, we chose a site located in the nearest intron. We constructed a donor template with the fragment containing the corrected point mutation as one of the homologous recombination arms flanking a PGK-Puro
Identifiants
pubmed: 33857832
pii: S1873-5061(21)00178-1
doi: 10.1016/j.scr.2021.102332
pmc: PMC8283763
mid: NIHMS1711907
pii:
doi:
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
102332Subventions
Organisme : NIAID NIH HHS
ID : R01 AI125650
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS075839
Pays : United States
Organisme : NINDS NIH HHS
ID : R03 NS107751
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.
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