Unpredicted central inversion in a sgRNA flanked by inverted repeats.

Abstract:

: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.

journal_name

Mol Biol Rep

authors

Wang G,Sukumar S

doi

10.1007/s11033-020-05524-1

subject

Has Abstract

pub_date

2020-08-01 00:00:00

pages

6375-6378

issue

8

eissn

0301-4851

issn

1573-4978

pii

10.1007/s11033-020-05524-1

journal_volume

47

pub_type

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