Large-scale transgenicDrosophilaresource collections for loss- and gain-of-function studies
| Autor: | Xia Wang, Jonathan Zirin, Ruibao Zhu, Ping Peng, Rong Tao, Shu Kondo, Dong Yan, Kuniaki Saito, Liz Perkins, Henna Ragoowansi, Jin Sun, Ethan Fenton, Ryan Colbeth, Benjamin Scott Ewen-Campen, Senait Efrem, Annette L. Parks, Limmond Ayisi, Sara VanNest, Donghui Yang-Zhou, Yuhao Qiu, Cooper Cavers, Norbert Perrimon, Aram Comjean, Jian-Quan Ni, Stephanie E. Mohr, Eric Vogt, Yanhui Hu, Jinchao Yu, Lu-Ping Liu, Da Shen, Christians Villalta, Yu Jia |
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| Rok vydání: | 2019 |
| Předmět: |
Computational biology
Investigations Biology Genome engineering Animals Genetically Modified 03 medical and health sciences 0302 clinical medicine Loss of Function Mutation RNA interference Databases Genetic Genetics Animals CRISPR Coding region Gene 030304 developmental biology Regulation of gene expression 0303 health sciences Cas9 fungi biology.organism_classification Drosophila melanogaster Gain of Function Mutation ComputingMethodologies_DOCUMENTANDTEXTPROCESSING ComputingMethodologies_GENERAL CRISPR-Cas Systems Genetic Engineering human activities Functional genomics 030217 neurology & neurosurgery |
| Zdroj: | Genetics |
| DOI: | 10.1101/852376 |
| Popis: | The Transgenic RNAi Project (TRiP), a Drosophila melanogaster functional genomics platform at Harvard Medical School, was initiated in 2008 to generate and distribute a genome-scale collection of RNA interference (RNAi) fly stocks. To date, it has generated >15,000 RNAi fly stocks. As this covers most Drosophila genes, we have largely transitioned to development of new resources based on CRISPR technology. Here, we present an update on our libraries of publicly available RNAi and CRISPR fly stocks, and focus on the TRiP-CRISPR overexpression (TRiP-OE) and TRiP-CRISPR knockout (TRiP-KO) collections. TRiP-OE stocks express single guide RNAs targeting upstream of a gene transcription start site. Gene activation is triggered by coexpression of catalytically dead Cas9 fused to an activator domain, either VP64-p65-Rta or Synergistic Activation Mediator. TRiP-KO stocks express one or two single guide RNAs targeting the coding sequence of a gene or genes. Cutting is triggered by coexpression of Cas9, allowing for generation of indels in both germline and somatic tissue. To date, we have generated >5000 TRiP-OE or TRiP-KO stocks for the community. These resources provide versatile, transformative tools for gene activation, gene repression, and genome engineering. |
| Databáze: | OpenAIRE |
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