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CRISPR/Cas9-mediated genome editing induces exon skipping by complete or stochastic altering splicing in the migratory locust
- Source :
- BMC Biotechnology, Vol 18, Iss 1, Pp 1-9 (2018)
- Publication Year :
- 2018
- Publisher :
- BMC, 2018.
-
Abstract
- Background The CRISPR/Cas9 system has been widely used to generate gene knockout/knockin models by inducing frameshift mutants in cell lines and organisms. Several recent studies have reported that such mutants can lead to in-frame exon skipping in cell lines. However, there was little research about post-transcriptional effect of CRISPR-mediated gene editing in vivo. Results We showed that frameshift indels also induced complete or stochastic exon skipping by deleting different regions to influence pre-mRNA splicing in vivo. In the migratory locust, the missing 55 bp at the boundary of intron 3 and exon 4 of an olfactory receptor gene, LmigOr35, resulted in complete exon 4 skipping, whereas the lacking 22 bp in exon 4 of LmigOr35 only resulted in stochastic exon 4 skipping. A single sgRNA induced small insertions or deletions at the boundary of intron and exon to disrupt the 3′ splicing site causing completely exon skipping, or alternatively induce small insertions or deletions in the exon to stochastic alter splicing causing the stochastic exon skipping. Conclusions These results indicated that complete or stochastic exon skipping could result from the CRISPR-mediated genome editing by deleting different regions of the gene. Although exon skipping caused by CRISPR-mediated editing was an unexpected outcome, this finding could be developed as a technology to investigate pre-mRNA splicing or to cure several human diseases caused by splicing mutations.
- Subjects :
- 0301 basic medicine
RNA Splicing
lcsh:Biotechnology
Grasshoppers
Biology
Gene editing
Receptors, Odorant
Frameshift mutation
03 medical and health sciences
Exon
Pre-mRNA splicing
Genome editing
INDEL Mutation
lcsh:TP248.13-248.65
CRISPR
Animals
Frameshift Mutation
Gene
CRISPR/Cas9
Sequence Deletion
Genetics
030102 biochemistry & molecular biology
Intron
Exons
Exon skipping
030104 developmental biology
Migratory locust
RNA splicing
Insect Proteins
Animal Migration
CRISPR-Cas Systems
Biotechnology
Subjects
Details
- Language :
- English
- ISSN :
- 14726750
- Volume :
- 18
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- BMC Biotechnology
- Accession number :
- edsair.doi.dedup.....5f069b6ae6d58ff488b3af6b32eb25bd