title: Chromosomal deletions and inversions mediated by TALENs and CRISPR/Cas in zebrafish.
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.
authors.affiliation: Key Laboratory of Cell Proliferation and Differentiation of Ministry of Education, College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: An
authors.initials: A
authors.lastname: Xiao
authors.forename: Zhanxiang
authors.initials: Z
authors.lastname: Wang
authors.forename: Yingying
authors.initials: Y
authors.lastname: Hu
authors.forename: Yingdan
authors.initials: Y
authors.lastname: Wu
authors.forename: Zhou
authors.initials: Z
authors.lastname: Luo
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Yang
authors.forename: Yao
authors.initials: Y
authors.lastname: Zu
authors.forename: Wenyuan
authors.initials: W
authors.lastname: Li
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Tong
authors.forename: Zuoyan
authors.initials: Z
authors.lastname: Zhu
authors.forename: Shuo
authors.initials: S
authors.lastname: Lin
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002872:Chromosome Deletion; D007446:Chromosome Inversion; D030541:Databases, Genetic; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods; D056426:Genetic Loci; D016678:Genome; D055029:Inverted Repeat Sequences; D035683:MicroRNAs / Q000235:genetics; D011506:Proteins / Q000235:genetics; D015027:Zebrafish / Q000235:genetics*; D058727:RNA, Small Untranslated
chemical_list: D035683:MicroRNAs; D011506:Proteins; D004706:Endodeoxyribonucleases; D058727:RNA, Small Untranslated