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. 2020 Aug;47(8):6375-6378.
doi: 10.1007/s11033-020-05524-1. Epub 2020 May 18.

Unpredicted central inversion in a sgRNA flanked by inverted repeats

Guannan Wang  1   2   3 , Saraswati Sukumar  4   5
Affiliations

Unpredicted central inversion in a sgRNA flanked by inverted repeats

Guannan Wang et al. Mol Biol Rep. 2020 Aug.

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.

Keywords: CRISPR/Cas9; Inversion; Inverted repeats; sgRNA.

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References

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