Functional Characterization of ASD-Associated EEF1A2 Mutations in Human Neurons
Abstract
We will determine how ASD-associated mutations in EEF1A2 impact protein synthesis and result in deficits in neuronal development and synaptic function in human neurons. Using human induced pluripotent stems cell (hiPSC)-derived neurons as a model, the CRISPR-Cas9 system will be utilized to recapitulate ASD-associated mutations in EEF1A2 observed in patients. The hiPSCs will then be differentiated into neurons using Neurogenin-2, a master transcription factor capable of inducing differentiation into excitatory neurons in under two weeks. Using this platform, the effect of ASD-associated mutations on neuronal function will be studied. First, we will determine the impact of three ASD-associated EEF1A2 mutations on protein synthesis in neurons, given the central role that EEF1A2 plays in protein synthesis. Moreover, we will perform ribosome profiling to determine the translatome and measure the elongation rate and translational efficiency associated with each EEF1A2 mutation. Then we will determine the impact of each EEF1A2 mutation on neuronal development and morphology, and synaptic function. The results of these studies will advance our understanding of the role translation elongation plays in neuronal development, and how its dysregulation leads to ASD-associated pathophysiologies.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2022
- Accession Number
- AD1169486
Entities
People
- Eric Klann
Organizations
- New York University