Unpredicted central inversion in a sgRNA flanked by inverted repeats

Guannan Wang, Saraswati Sukumar

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)6375-6378
Number of pages4
JournalMolecular Biology Reports
Volume47
Issue number8
DOIs
StatePublished - Aug 1 2020

Keywords

  • CRISPR/Cas9
  • Inversion
  • Inverted repeats
  • sgRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'Unpredicted central inversion in a sgRNA flanked by inverted repeats'. Together they form a unique fingerprint.

Cite this