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题名: Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs
作者: Liu, Yun1, 4; Luo, Daji2, 3; Zhao, Hui1, 4; Zhu, Zuoyan2; Hu, Wei2; Cheng, Christopher H. K.1, 4
通讯作者: Hu, W (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan, Peoples R China.
关键词: ZINC-FINGER NUCLEASES ; EFFECTOR NUCLEASES ; CHROMOSOMAL DELETIONS ; EMBRYONIC-DEVELOPMENT ; MICRORNA EXPRESSION ; TRANSCRIPTION ; CELLS ; KNOCKOUT ; ORGANIZATION ; RECOGNITION
刊名: PLOS ONE
发表日期: 2013-10-10
DOI: 10.1371/journal.pone.0076387
卷: 8, 期:10, 页:e76387
收录类别: SCI
文章类型: Article
部门归属: [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
WOS标题词: Science & Technology
资助者: Research Grant Council of Hong Kong [462312]; National Natural Science Foundation of China [31172394, 30900853, 30930069]; Chinese Academy of Sciences [KSCX2-EW-N-004]
类目[WOS]: Multidisciplinary Sciences
研究领域[WOS]: Science & Technology - Other Topics
摘要: 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.
英文摘要: 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.
关键词[WOS]: ZINC-FINGER NUCLEASES ; CHROMOSOMAL DELETIONS ; EFFECTOR NUCLEASES ; EMBRYONIC-DEVELOPMENT ; MICRORNA EXPRESSION ; TRANSCRIPTION ; CELLS ; KNOCKOUT ; ORGANIZATION ; RECOGNITION
语种: 英语
WOS记录号: WOS:000325814200038
ISSN号: 1932-6203
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/19571
Appears in Collections:鱼类生物学及渔业生物技术研究中心_期刊论文

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作者单位: 1.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:
Liu, Yun; Luo, Daji; Zhao, Hui; Zhu, Zuoyan; Hu, Wei; Cheng, Christopher H. K..Inheritable and Precise Large Genomic Deletions of Non-Coding RNA Genes in Zebrafish Using TALENs,PLOS ONE,2013,8(10):e76387
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