Optical pooled CRISPR screen in iPSC-derived human ioMicroglia using Element Biosciences AVITI24 system

Targets with human genetic evidence are twice as likely to succeed in clinical trials and gain drug approval compared to those without (Nelson et al., Nature Genetics, 2015). Identifying and validating such targets requires systematic approaches that can connect genetic perturbations to complex cellular phenotypes in physiologically relevant models.

You will learn:

• How CRISPRko-Ready ioMicroglia combined with the Element Biosciences AVITI24 system enable simultaneous in situ detection of sgRNA identity, protein abundance and cell morphology in a single experiment

• How optical sgRNA detection recovered distinct perturbation identities across spatially distributed, morphologically heterogeneous cells, with library representation and target coverage comparable to conventional pooled CRISPR screens and a median of 109 cells per perturbation in each polarisation state

• How protein and morphology profiling discriminates M0, M1- and M2-like microglia states, and how HDAC3, DOT1L and PRMT6 knockouts shift microglia away from the M1 state, consistent with an essential role for these genes in maintaining it