Unpredicted central inversion in a sgRNA flanked by inverted repeats.
CRISPR/Cas9
Inversion
Inverted repeats
sgRNA
Journal
Molecular biology reports
ISSN: 1573-4978
Titre abrégé: Mol Biol Rep
Pays: Netherlands
ID NLM: 0403234
Informations de publication
Date de publication:
Aug 2020
Aug 2020
Historique:
received:
23
10
2019
accepted:
14
05
2020
pubmed:
20
5
2020
medline:
27
5
2021
entrez:
20
5
2020
Statut:
ppublish
Résumé
In genome engineering, sgRNAs define the genomic target to be modified in CRISPR/Cas9 system. Either for single gene editing or genome-wide screens, sgRNAs are cloned into plasmid vectors. During our performance of CRISPR/Cas9 gene knock out, we found that the central part of a sgRNA was inverted after transformation into Escherichia coli. Interestingly, the inverted portion was found to be flanked by inverted repeats, and sealing of nicks inside the plasmid could correct the inversion. This type of sgRNA recombination completely changed its original sequence and should be noted during sgRNA design and performance of CRISPR/Cas9.
Identifiants
pubmed: 32424520
doi: 10.1007/s11033-020-05524-1
pii: 10.1007/s11033-020-05524-1
doi:
Substances chimiques
HOXB13 protein, human
0
Homeodomain Proteins
0
RNA, Guide
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
6375-6378Références
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