Multiplex analysis of variant effects for SLC2A1 gene
Awardee: Christina Gurnett
Institution: Washington University, St Louis
Grant Amount: $64,465
Funding Period: February 1, 2022 - January 31, 2023
Summary:
The goal of this research is to quantitatively determine the functional impact of all possible genetic variants in SLC2A1 and construct algorithms to accurately predict disease onset and severity correlating through calibration with known pathogenic and benign variants. Toward this goal, we propose to employ a high-throughput framework to assess the functional impact of genetic variants in the SLC2A1. We will introduce hundreds of SLC2A1 variants individually into the haploid cell line (HAP1) via multiplex homology-directed-repair (HDR) using CRISPR and a donor library, so that each cell obtains a single variant knocked into the endogenous SLC2A1 gene. Damaging variants that completely disrupt SLC2A1 function (and glucose uptake) will result in impaired cell growth and will drop out of a population of cells sequenced at different time points. We have preliminary data demonstrating the utility of this approach to quantify the functional effects of 15 SLC2A1 variants. We now propose to scale this assay to generate comprehensive, quantitative functional data for the entire SLC2A1 coding region.