Transgenic RNAi Project (TRiP) Platform at the DRSC/TRiP Functional Genomics Resources
Grant Numbers: R24OD030002, R24OD031952, R24OD035556
Research Emphasis/Objectives
The Transgenic RNAi Project (TRiP) is an in vivo functional genetics platform that has generated more than 20,000 short hairpin RNAs (shRNA) and single-guide RNAs (sgRNA) fly stocks for the research community. These resources, which are distributed by the Bloomington Drosophila Stock Center (BDSC), provide powerful, versatile, and transformative tools for gene knockdown, knockout, and activation. Researchers have used these fly stocks extensively to model human diseases associated with altered gene expression.
Current Research
To complement the shRNA and sgRNA collections, new in vivo resources were generated to support three aims. First, the resources enabled combinatorial studies using multiple binary expression systems to study signaling between different tissues and cell types. Focusing on genes with well-characterized GAL4 expression patterns, a set of more than 40 tissue-specific LexA-GAD and QF2 insertions by CRISPR knock-in were generated. Second, the resource enabled the validation of scRNA gene clusters using split-Gal4 lines. Using an algorithm that determines the smallest unique set of marker genes that define a cluster, several hundred highly specific Gal80-repressible split-Gal4 knock-in lines were generated. Last, the resource supported the detection of fly proteins through the insertion of a NanoTag epitope at the C-terminus of more than 300 high-confidence Drosophila orthologs of human mitochondrial disease genes that can then be recognized by an existing high-affinity nanobody against the NanoTag. Together, these new in vivo resources will dramatically expand the scope of reagents available to researchers who are using the fly model to understand the etiology of human diseases.
Services Provided
The TRiP supports the production of fly stocks with shRNAs for gene knockdown; sgRNAs targeting gene-coding sequence for gene knockout; sgRNAs targeting upstream gene regions, such that they are useful for CRISPR overexpression; and CRISPR knock-in lines for LexA/QF, Split-Gal4, and endogenous epitope tags. Immediately following production and validation, the fly stocks are shared with the BDSC for distribution to the community.
Contact Information
Jonathan Zirin, Ph.D.
Assistant Director of DRSC/TRiP Functional Genomics Resources
Harvard Medical School
New Research Building, Room 336
77 Avenue Louis Pasteur
Boston, MA 02115
Email: [email protected]
Principal Investigator
Norbert Perrimon
Harvard Medical School
Phone: 617-432-7672
Email: [email protected]