Award
National Institute of Neurological Disorders and Stroke 4DP2NS131566-03
Methods to Rapidly Explore Combinatorial Diversity and their Application to CRISPR-Cas9 Systems
Recipient
University of California, San Diego
Award Amount
$955,050.00
Ceiling
$955,050.00
Awarded
January 28, 2026
Identifier
4DP2NS131566-03
The award funds the development of BaCES, a high-throughput method for creating and testing combinatorial protein variants, specifically enhanced Cas9 transcriptional regulators, to enable in vivo genetic screens in neurodegeneration research.
Description
This project introduces BaCES, a method enabling the simultaneous assembly and parallel testing of tens of thousands of combinatorial protein variants. The objective is to use BaCES to create a new generation of enhanced Cas9-based transcriptional regulators, combined with a novel paradigm to probe gene function within in vivo contexts. The project aims to generate and quantify 27,000 Cas9 activators and repressors, validate highly-potent Cas9 transcriptional modulators across targets and cell types, and perform in vivo genetic screens to uncover regulators of neuronal survival in a mouse model of Parkinson’s Disease. This innovative approach will facilitate rapid searching through combinatorial protein space and enable in vivo CRISPR screens within complex environments, advancing understanding of neuronal tolerance to neurodegenerative insults.