首页    期刊浏览 2024年11月27日 星期三
登录注册

文章基本信息

  • 标题:Dual sgRNA-directed gene knockout using CRISPR/Cas9 technology in Caenorhabditis elegans
  • 本地全文:下载
  • 作者:Xiangyang Chen ; Fei Xu ; Chengming Zhu
  • 期刊名称:Scientific Reports
  • 电子版ISSN:2045-2322
  • 出版年度:2014
  • 卷号:4
  • DOI:10.1038/srep07581
  • 出版社:Springer Nature
  • 摘要:The CRISPR RNA-guided Cas9 nuclease gene-targeting system has been successfully used for genome editing in a variety of organisms. Here, we report the use of dual sgRNA-guided Cas9 nuclease to generate knockout mutants of protein coding genes, noncoding genes, and repetitive sequences in C. elegans . Co-injection of C. elegans with dual sgRNAs results in the removal of the interval between two sgRNAs and the loss-of-function phenotype of targeted genes. We sought to determine how large an interval can be eliminated and found that at least a 24 kb chromosome segment can be deleted using this dual sgRNA/Cas9 strategy. The deletion of large chromosome segments facilitates mutant screening by PCR and agarose electrophoresis. Thus, the use of the CRISPR/Cas9 system in combination with dual sgRNAs provides a powerful platform with which to easily generate gene knockout mutants in C. elegans . Our data also suggest that encoding multiple sgRNA sequences into a single CRISPR array to simultaneously edit several sites within the genome may cause the off-target deletion of chromosome sequences.
国家哲学社会科学文献中心版权所有