Forward Genetic Screen to Identify Novel Therapeutic Entry Points of an Autism Spectrum Disorder
Technical Report,01 Aug 2018,31 Jul 2019
Baylor College of Medicine Houston United States
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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 EGFPShank3. The positive hits from the kinasephosphatase sub-library were then tested in a secondary screen with siRNA depletion of those hits and measurement of EGFPShank3 abundance by flow cytometry. The two most robustly increased gene from the kinasephosphatase 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.
- Medicine and Medical Research
- Genetic Engineering and Molecular Biology