PUBLICATION

Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish

Authors
Xiao, A., Wang, Z., Hu, Y., Wu, Y., Luo, Z., Yang, Z., Zu, Y., Li, W., Huang, P., Tong, X., Zhu, Z., Lin, S., and Zhang, B.
ID
ZDB-PUB-130709-35
Date
2013
Source
Nucleic acids research   41(14): e141 (Journal)
Registered Authors
Huang, Peng, Hu, Yingying, Lin, Shuo, Li, Wenyuan, Luo, Zhou, Tong, Xiangjun, Wang, Zhanxiang, Xiao, An, Zhang, Bo, Zu, Yao
Keywords
none
MeSH Terms
  • Animals
  • Chromosome Deletion*
  • Chromosome Inversion*
  • Databases, Genetic
  • Endodeoxyribonucleases/metabolism*
  • Genetic Engineering/methods
  • Genetic Loci
  • Genome
  • Inverted Repeat Sequences*
  • MicroRNAs/genetics
  • Proteins/genetics
  • RNA, Guide/chemistry
  • Zebrafish/genetics*
PubMed
23748566 Full text @ Nucleic Acids Res.
Abstract

Customized TALENs and Cas9/gRNAs have been used for targeted mutagenesis in zebrafish to induce indels into protein-coding genes. However, indels are usually not sufficient to disrupt the function of non-coding genes, gene clusters or regulatory sequences, whereas large genomic deletions or inversions are more desirable for this purpose. By injecting two pairs of TALEN mRNAs or two gRNAs together with Cas9 mRNA targeting distal DNA sites of the same chromosome, we obtained predictable genomic deletions or inversions with sizes ranging from several hundred bases to nearly 1 Mb. We have successfully achieved this type of modifications for 11 chromosomal loci by TALENs and 2 by Cas9/gRNAs with different combinations of gRNA pairs, including clusters of miRNA and protein-coding genes. Seven of eight TALEN-targeted lines transmitted the deletions and one transmitted the inversion through germ line. Our findings indicate that both TALENs and Cas9/gRNAs can be used as an efficient tool to engineer genomes to achieve large deletions or inversions, including fragments covering multiple genes and non-coding sequences. To facilitate the analyses and application of existing ZFN, TALEN and CRISPR/Cas data, we have updated our EENdb database to provide a chromosomal view of all reported engineered endonucleases targeting human and zebrafish genomes.

Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes