CRISPR/Cas9-mediated genome editing induces gene knockdown by altering the pre-mRNA splicing in mice.

Abstract:

BACKGROUND:Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9 (CRISPR/Cas9) has been wildly used to generate gene knockout models through inducing indels causing frame-shift. However, there are few studies concerning the post-transcript effects caused by CRISPR-mediated genome editing. RESULTS:In the present study, we showed that gene knockdown model also could be generated using CRISPR-mediated gene editing by disrupting the boundary of exon and intron in mice (C57BL/6 J). CRISPR induced indel at the boundary of exon and intron (5' splice site) caused alternative splicing and produced multiple different mRNAs, most of these mRNAs introduced premature termination codon causing down expression of the gene. CONCLUSIONS:These results showed that alternative splicing mutants were able to generate through CRISPR-mediated genome editing by deleting the boundary of exon and intron causing disruption of 5' splice site. Although alternative splicing was an unexpected outcome, this finding could be developed as a technology to generate gene knockdown models or to investigate pre-mRNA splicing.

journal_name

BMC Biotechnol

journal_title

BMC biotechnology

authors

Tang JX,Chen D,Deng SL,Li J,Li Y,Fu Z,Wang XX,Zhang Y,Chen SR,Liu YX

doi

10.1186/s12896-018-0472-8

subject

Has Abstract

pub_date

2018-10-03 00:00:00

pages

61

issue

1

issn

1472-6750

pii

10.1186/s12896-018-0472-8

journal_volume

18

pub_type

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