Unknown

Dataset Information

0

Optimized gene editing technology for Drosophila melanogaster using germ line-specific Cas9.


ABSTRACT: 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.

SUBMITTER: Ren X 

PROVIDER: S-EPMC3839733 | biostudies-other | 2013 Nov

REPOSITORIES: biostudies-other

altmetric image

Publications

Optimized gene editing technology for Drosophila melanogaster using germ line-specific Cas9.

Ren Xingjie X   Sun Jin J   Housden Benjamin E BE   Hu Yanhui Y   Roesel Charles C   Lin Shuailiang S   Liu Lu-Ping LP   Yang Zhihao Z   Mao Decai D   Sun Lingzhu L   Wu Qujie Q   Ji Jun-Yuan JY   Xi Jianzhong J   Mohr Stephanie E SE   Xu Jiang J   Perrimon Norbert N   Ni Jian-Quan JQ  

Proceedings of the National Academy of Sciences of the United States of America 20131104 47


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 in  ...[more]

Similar Datasets

| S-EPMC1805588 | biostudies-literature
| S-EPMC8459378 | biostudies-literature
| S-EPMC8715323 | biostudies-literature
| S-EPMC1205432 | biostudies-other
| S-EPMC6429391 | biostudies-literature
| S-EPMC9210308 | biostudies-literature
| S-EPMC6423289 | biostudies-literature
| S-EPMC5706864 | biostudies-literature
| S-EPMC444866 | biostudies-other
| S-EPMC1871851 | biostudies-other