期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 110, 期 47, 页码 19012-19017出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1318481110
关键词
nanos-Cas9; HRMA
资金
- Tsinghua-Peking Center for Life Sciences
- Ministry of Science and Technology of China 973 Program [2013CB835100]
- PhD Programs Foundation of the Chinese Ministry of Education [20121018577, 20120002110056]
- National Institutes of Health, National Institute of General Medical Sciences [R01 GM084947, R01 GM067761]
The ability to engineer genomes in a specific, systematic, and cost-effective way is critical for functional genomic studies. Recent advances using the CRISPR-associated single-guide RNA system (Cas9/sgRNA) illustrate the potential of this simple system for genome engineering in a number of organisms. Here we report an effective and inexpensive method for genome DNA editing in Drosophila melanogaster whereby plasmid DNAs encoding short sgRNAs under the control of the U6b promoter are injected into transgenic flies in which Cas9 is specifically expressed in the germ line via the nanos promoter. We evaluate the off-targets associated with the method and establish a Web-based resource, along with a searchable, genome-wide database of predicted sgRNAs appropriate for genome engineering in flies. Finally, we discuss the advantages of our method in comparison with other recently published approaches.
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。
推荐
暂无数据