التفاصيل البيبلوغرافية
| العنوان: |
Efficient genome-wide mutagenesis of zebrafish genes by retroviral insertions. |
| المؤلفون: |
Dongmei Wang, Li-en Jao, Naizhong Zheng, Kyle Dolan, Jessica Ivey, Seth Zonies, Xiaolin Wu, Kangmai Wu, Hongbo Yang, Qingchao Meng, Zuoyan Zhu, Bo Zhang, Shuo Lin, Shawn M. Burgess |
| المصدر: |
Proceedings of the National Academy of Sciences of the United States of America; 7/24/2007, Vol. 104 Issue 30, p12428-12433, 6p, 2 Charts, 4 Graphs |
| مصطلحات موضوعية: |
HEREDITY, ZEBRA danio, GENETICS, GENES, ONCOGENIC viruses, MESSENGER RNA |
| مستخلص: |
Using a combination of techniques we developed, we infected zebrafish embryos using pseudotyped retroviruses and mapped the genomic locations of the proviral integrations in the F1 off- spring of the infected fish. From F1 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 ∼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 F1 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 F1 fish, it is now possible to create a stable resource that contains mutations in every known zebrafish gene. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
