4.8 Article

Directed, efficient, and versatile modifications of the Drosophila genome by genomic engineering

出版社

NATL ACAD SCIENCES
DOI: 10.1073/pnas.0900641106

关键词

cell polarity; ends-out targeting; homologous recombination; phiC31 integrase

资金

  1. University of Pittsburgh Medical School
  2. National Center for Research Resources, National Institutes of Health [1R21RR024869]

向作者/读者索取更多资源

With the completion of genome sequences of major model organisms, increasingly sophisticated genetic tools are necessary for investigating the complex and coordinated functions of genes. Here we describe a genetic manipulation system termed genomic engineering'' in Drosophila. Genomic engineering is a 2-step process that combines the ends-out (replacement) gene targeting with phage integrase phi C31-mediated DNA integration. First, through an improved and modified gene targeting method, a founder knockout line is generated by deleting the target gene and replacing it with an integration site of phi C31. Second, DNA integration by phi C31 is used to reintroduce modified target-gene DNA into the native locus in the founder knock-out line. Genomic engineering permits directed and highly efficient modifications of a chosen genomic locus into virtually any desired mutant allele. We have successfully applied the genomic engineering scheme on 6 different genes and have generated at their loci more than 70 unique alleles.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.8
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据