期刊
BIO-PROTOCOL
卷 9, 期 2, 页码 -出版社
BIO-PROTOCOL
DOI: 10.21769/BioProtoc.3141
关键词
Genome editing; CRISPR; Cas9; Drosophila; Heritable mutations
类别
资金
- NIH [R01GM084947, R24OD021997]
- NIH's Ruth L. Kirschstein National Research Service Award from the NIH General Medical Sciences Division [F32GM113395]
- National Key Technology Research and Development Program of the Ministry of Science and Technology of the People's Republic of China [2015BAI09B03, 2016YFE0113700]
- National Natural Science Foundation of China [31571320]
In recent years, great progress has been made in the research of genome editing systems, one of which is the CRISPR-Cas9 system, a powerful technology that is applied to edit animal genome. Here, we describe a CRISPR-Cas9 mediated mutation protocol for efficiently and specifically editing genes in Drosophila. In this optimized system, the mutant progeny can be generated by only injecting a DNA plasmid encoding synthetic guide RNA (sgRNA) under the control of the U6b promoter into transgenic fly embryos in which Cas9 is specifically expressed in the progenitor cells, thus the gene of interest can be edited by the CRISPR in germ cells, with high rate of heritable mutations and few side effects.
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