Efficient genome-wide mutagenesis of zebrafish genes by retroviral   insertions

doi:10.1073/pnas.0705502104

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	Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions
	Wang, Dongmei ; Jao, Li-En ; Zheng, Naizhong ; Dolan, Kyle ; Ivey, Jessica ; Zonles, Seth ; Wul, Xiaolin ; Wu, Kangmai ; Yang, Hongbo ; Meng, Qingchao ; Zhu, Zuoyan ; Zhang, Bo ; Lin, Shuo ; Burgess, Shawn M.
刊名	proceedings of the national academy of sciences of the united states of america
	2007
关键词	genetics retrovirus GERM-LINE TRANSMISSION FUNCTIONAL GENOMICS POSITIONAL CLONING CELL-LINES HIGH-TITER VECTOR INTEGRATION MUTATIONS MOUSE
DOI	10.1073/pnas.0705502104
英文摘要	Using a combination of techniques we developed, we infected zebrafish embryos using pseuclotyped retroviruses and mapped the genomic locations of the proviral integrations in the F, offspring of the infected fish. From F, fish, we obtained 2,045 sequences representing 933 unique retroviral integrations. A total of 599 were mappable to the current genomic assembly (Zv6), and 233 of the integrations landed within genes. By inbreeding fish carrying proviral integrations in 25 different genes, we were able to demonstrate that in approximate to 50% of the gene "hits," the mRNA transcript levels were reduced by >= 70%, with the highest probability for mutation occurring if the integration was in an exon or first intron. Based on these data, the mutagenic frequency for the retrovirus is nearly one in five integrations. In addition, a strong mutagenic effect is seen when murine leukemia virus integrates specifically in the first intron of genes but not in other introns. Three of 19 gene inactivation events had embryonic defects. Using the strategy we outlined, it is possible to identify 1 mutagenic event for every 30 sequencing reactions done on the F, fish. This is a 20- to 30-fold increase in efficiency when compared with the current resequencing approach [targeting induced local lesions in genomes (TILLING)] used in zebrafish for identifying mutations in genes. Combining this increase in efficiency with cryopreservation of sperm samples from the F, fish, it is now possible to create a stable resource that contains mutations in every known zebrafish gene.; Multidisciplinary Sciences; SCI(E); PubMed; 61; ARTICLE; 30; 12428-12433; 104
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/345387]
专题	生命科学学院
推荐引用方式 GB/T 7714	Wang, Dongmei,Jao, Li-En,Zheng, Naizhong,et al. Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions[J]. proceedings of the national academy of sciences of the united states of america,2007.
APA	Wang, Dongmei.,Jao, Li-En.,Zheng, Naizhong.,Dolan, Kyle.,Ivey, Jessica.,...&Burgess, Shawn M..(2007).Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions.proceedings of the national academy of sciences of the united states of america.
MLA	Wang, Dongmei,et al."Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions".proceedings of the national academy of sciences of the united states of america (2007).