Successful delivery of large-size CRISPR/Cas9 vectors in hard-to-transfect human cells using small plasmids.
Journal
Communications biology
ISSN: 2399-3642
Titre abrégé: Commun Biol
Pays: England
ID NLM: 101719179
Informations de publication
Date de publication:
19 06 2020
19 06 2020
Historique:
received:
15
11
2019
accepted:
29
05
2020
entrez:
21
6
2020
pubmed:
21
6
2020
medline:
24
6
2021
Statut:
epublish
Résumé
With the rise of new powerful genome engineering technologies, such as CRISPR/Cas9, cell models can be engineered effectively to accelerate basic and disease research. The most critical step in this procedure is the efficient delivery of foreign nucleic acids into cells by cellular transfection. Since the vectors encoding the components necessary for CRISPR/Cas genome engineering are always large (9-19 kb), they result in low transfection efficiency and cell viability, and thus subsequent selection or purification of positive cells is required. To overcome those obstacles, we here show a non-toxic and non-viral delivery method that increases transfection efficiency (up to 40-fold) and cell viability (up to 6-fold) in a number of hard-to-transfect human cancer cell lines and primary blood cells. At its core, the technique is based on adding exogenous small plasmids of a defined size to the transfection mixture.
Identifiants
pubmed: 32561814
doi: 10.1038/s42003-020-1045-7
pii: 10.1038/s42003-020-1045-7
pmc: PMC7305135
doi:
Substances chimiques
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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