PUBLICATION
The novel insight into the outcomes of CRISPR/Cas9 editing intra- and inter-species
- Authors
- Chang, J., Chen, X., Zhang, T., Wang, R., Wang, A., Lan, X., Zhou, Y., Ma, S., Xia, Q.
- ID
- ZDB-PUB-200712-9
- Date
- 2020
- Source
- International journal of biological macromolecules 163: 711-717 (Journal)
- Registered Authors
- Keywords
- CRISPR/Cas9, DNA manipulation, DNA repair, Deep sequencing, Genome editing, Indel patterns
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems*
- Cell Line
- Gene Editing*
- Genetic Vectors
- High-Throughput Nucleotide Sequencing
- Humans
- INDEL Mutation
- Mice
- Mutation
- RNA, Guide, Kinetoplastida
- Species Specificity
- Zebrafish
- PubMed
- 32652159 Full text @ Int. J. Biol. Macromol.
Citation
Chang, J., Chen, X., Zhang, T., Wang, R., Wang, A., Lan, X., Zhou, Y., Ma, S., Xia, Q. (2020) The novel insight into the outcomes of CRISPR/Cas9 editing intra- and inter-species. International journal of biological macromolecules. 163:711-717.
Abstract
The CRISPR/Cas (clustered regularly interspaced short palindromic repeat technology/CRISPR-associated protein) is a widely used and powerful research tool in biosciences and a promising therapeutic agent for treating genetic diseases. Mutations induced by Cas9 are generally considered stochastic and unpredictable, thus hindering its applications where precise genetic alternations are required. Here, through deep sequencing and analysis of genome editing outcomes of multiple sites in four distinct species, we found that Cas9-induced mutations are coincident in mutation types but are significantly different in indel patterns among species. In human and mouse cells, indels were almost evenly distributed at both ends of the cleavage sites. However, the indels mainly appeared at the upstream of cleavage sites in Bombyx mori, while they predominantly occurred downstream of the cleavage sites in the zebrafish Danio rerio. We also found that within a species, indel patterns are sequence dependent, wherein deletions between two adjacent micro-homology sequences were the most frequently observed mutations in the repair spectrum. These results suggested the species differences in DNA repair processes during Cas9-induced gene editing, and the important role of sequence structure at the target site in predicting the gene editing outcome.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping