Forward Genetic Screen to Identify Novel Therapeutic Entry Points of an Autism Spectrum Disorder
Abstract
During the reporting period, the PCR libraries from the primary screen were sequenced. The sequencing data sets then underwent quality control. Bioinformatics of the sequencing data identified those genes with significantly increased abundance of EGFP:Shank3. The positive hits from the kinase/phosphatase sub-library were then tested in a secondary screen with siRNA depletion of those hits and measurement of EGFP:Shank3 abundance by flow cytometry. The two most robustly increased gene from the kinase/phosphatase sub-library are now undergoing validation by Western blotting. These same two candidate hits will be tested for Shank3 protein regulation by knockdown in primary mouse neurons. We have already discovered that the protein complex for one of these hits interacts with Shank3 in vivo.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2019
- Accession Number
- AD1094712
Entities
People
- Jimmy Jr L. Holder
Organizations
- Baylor College of Medicine