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 Repjournal_title
Molecular biology reportsauthors
Wang G,Sukumar Sdoi
10.1007/s11033-020-05524-1subject
Has Abstractpub_date
2020-08-01 00:00:00pages
6375-6378issue
8eissn
0301-4851issn
1573-4978pii
10.1007/s11033-020-05524-1journal_volume
47pub_type
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