A transient CRISPR/Cas9 expression system for genome editing in Trypanosoma brucei.
CRISPR/Cas9
Transient transfection
Trypanosome
Journal
BMC research notes
ISSN: 1756-0500
Titre abrégé: BMC Res Notes
Pays: England
ID NLM: 101462768
Informations de publication
Date de publication:
03 Jun 2020
03 Jun 2020
Historique:
received:
04
04
2020
accepted:
13
05
2020
entrez:
5
6
2020
pubmed:
5
6
2020
medline:
5
3
2021
Statut:
epublish
Résumé
Generation of knockouts and in situ tagging of genes in Trypanosoma brucei has been greatly facilitated by using CRISPR/Cas9 as a genome editing tool. To date, this has entailed using a limited number of cell lines that are stably transformed to express Cas9 and T7 RNA polymerase (T7RNAP). It would be desirable, however, to be able to use CRISPR/Cas9 for any trypanosome cell line. We describe a sequential transfection expression system that enables transient expression of the two proteins, followed by delivery of PCR products for gRNAs and repair templates. This procedure can be used for genome editing without the need for stable integration of the Cas9 and T7RNAP genes.
Identifiants
pubmed: 32493474
doi: 10.1186/s13104-020-05089-z
pii: 10.1186/s13104-020-05089-z
pmc: PMC7268226
doi:
Substances chimiques
Viral Proteins
0
bacteriophage T7 RNA polymerase
EC 2.7.7.-
DNA-Directed RNA Polymerases
EC 2.7.7.6
RNA, Guide, Kinetoplastida
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
268Subventions
Organisme : Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
ID : 310030_184669
Organisme : Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
ID : 31003A_166427
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