IHB OpenIR  > 鱼类生物学及渔业生物技术研究中心  > 期刊论文
Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs
Liu, Yun1,4; Luo, Daji2,3; Zhao, Hui1,4; Zhu, Zuoyan2; Hu, Wei2; Cheng, Christopher H. K.1,4; Hu, W (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China.
2013-10-10
Source PublicationPLOS ONE
ISSN1932-6203
Volume8Issue:10Pages:e76387
AbstractTranscription activator-like effector nucleases (TALENs) have so far been applied to disrupt protein-coding genes which constitute only 2-3% of the genome in animals. The majority (70-90%) of the animal genome is actually transcribed as non-coding RNAs (ncRNAs), yet the lack of efficient tools to knockout ncRNA genes hinders studies on their in vivo functions. Here we have developed novel strategies using TALENs to achieve precise and inheritable large genomic deletions and knockout of ncRNA genes in zebrafish. We have demonstrated that individual miRNA genes could be disrupted using one pair of TALENs, whereas large microRNA (miRNA) gene clusters and long non-coding RNA (lncRNA) genes could be precisely deleted using two pairs of TALENs. We have generated large genomic deletions of two miRNA clusters (the 1.2 kb miR-17-92 cluster and the 79.8 kb miR-430 cluster) and one long non-coding RNA (lncRNA) gene (the 9.0 kb malat1), and the deletions are transmitted through the germline. Taken together, our results establish TALENs as a robust tool to engineer large genomic deletions and knockout of ncRNA genes, thus opening up new avenues in the application of TALENs to study the genome in vivo.; Transcription activator-like effector nucleases (TALENs) have so far been applied to disrupt protein-coding genes which constitute only 2-3% of the genome in animals. The majority (70-90%) of the animal genome is actually transcribed as non-coding RNAs (ncRNAs), yet the lack of efficient tools to knockout ncRNA genes hinders studies on their in vivo functions. Here we have developed novel strategies using TALENs to achieve precise and inheritable large genomic deletions and knockout of ncRNA genes in zebrafish. We have demonstrated that individual miRNA genes could be disrupted using one pair of TALENs, whereas large microRNA (miRNA) gene clusters and long non-coding RNA (lncRNA) genes could be precisely deleted using two pairs of TALENs. We have generated large genomic deletions of two miRNA clusters (the 1.2 kb miR-17-92 cluster and the 79.8 kb miR-430 cluster) and one long non-coding RNA (lncRNA) gene (the 9.0 kb malat1), and the deletions are transmitted through the germline. Taken together, our results establish TALENs as a robust tool to engineer large genomic deletions and knockout of ncRNA genes, thus opening up new avenues in the application of TALENs to study the genome in vivo.
SubtypeArticle
KeywordZinc-finger Nucleases Effector Nucleases Chromosomal Deletions Embryonic-development Microrna Expression Transcription Cells Knockout Organization Recognition
Department[Liu, Yun ; Zhao, Hui ; Cheng, Christopher H. K.] Chinese Univ Hong Kong, Sch Biomed Sci, Hong Kong, Hong Kong, Peoples R China ; [Luo, Daji ; Zhu, Zuoyan ; Hu, Wei] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China ; [Luo, Daji] Wuhan Univ, Sch Basic Med Sci, Dept Genet, Wuhan 430072, Peoples R China ; [Liu, Yun ; Zhao, Hui ; Cheng, Christopher H. K.] Chinese Univ Hong Kong, Shenzhen Res Inst, Sch Biomed Sci, Core Lab, Shenzhen, Peoples R China
DOI10.1371/journal.pone.0076387
WOS HeadingsScience & Technology
Funding OrganizationResearch Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004]
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
WOS SubjectMultidisciplinary Sciences
WOS IDWOS:000325814200038
WOS KeywordZINC-FINGER NUCLEASES ; CHROMOSOMAL DELETIONS ; EFFECTOR NUCLEASES ; EMBRYONIC-DEVELOPMENT ; MICRORNA EXPRESSION ; TRANSCRIPTION ; CELLS ; KNOCKOUT ; ORGANIZATION ; RECOGNITION
Funding OrganizationResearch Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004] ; Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004]
Citation statistics
Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/19571
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorHu, W (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China.
Affiliation1.Chinese Univ Hong Kong, Sch Biomed Sci, Hong Kong, Hong Kong, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China
3.Wuhan Univ, Sch Basic Med Sci, Dept Genet, Wuhan 430072, Peoples R China
4.Chinese Univ Hong Kong, Shenzhen Res Inst, Sch Biomed Sci, Core Lab, Shenzhen, Peoples R China
Recommended Citation
GB/T 7714
Liu, Yun,Luo, Daji,Zhao, Hui,et al. Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs[J]. PLOS ONE,2013,8(10):e76387.
APA Liu, Yun.,Luo, Daji.,Zhao, Hui.,Zhu, Zuoyan.,Hu, Wei.,...&Hu, W .(2013).Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs.PLOS ONE,8(10),e76387.
MLA Liu, Yun,et al."Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs".PLOS ONE 8.10(2013):e76387.
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