Forward Genetic Screen to Identify Novel Therapeutic Entry Points of an Autism Spectrum Disorder

Abstract

During the life of this grant, we have successfully performed a cell based CRISPR/Cas9screen to identify regulators of Shank3 protein stability from three sub-libraries,kinases/phosphatases, G-protein coupled receptors and ubiquitin-related proteins. We haveperformed a secondary screen with siRNA knockdown of the most promising hits from theprimary screen. Several of these hits were validated in the secondary screen. For one ofthe kinase hits, a small molecule inhibitor of the hit was found to also increase Shank3abundance. Thus, we have successfully proven the validity of the approach for identifyingregulators of protein stability for Shank3. We will next take our most promising validatedhits to a mouse in vivo system to confirm rescue of behavioral and molecular phenotypes.

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Document Details

Document Type
Technical Report
Publication Date
Nov 01, 2020
Accession Number
AD1125013

Entities

People

  • Jimmy L. Holder

Organizations

  • Baylor College of Medicine

Tags

DTIC Thesaurus Topics

  • Abstracts
  • Autism
  • Biomedical Research
  • Biomolecules
  • Cell Line
  • Cells
  • Cultured Cells
  • Department Of Defense
  • Diseases And Disorders
  • Factor Analysis
  • Information Operations
  • Inhibitors
  • Kinases
  • Maryland
  • Mass Spectrometry
  • Molecules
  • Phenotypes
  • Phosphorylation
  • Proteins
  • Quality Control
  • Regulators
  • Small Molecules
  • Spectra
  • Statistical Analysis
  • Therapy
  • Validation

Fields of Study

  • Biology

Readers

  • Molecular Genetics
  • Molecular and Cellular Biology

Technology Areas

  • Biotechnology