Single-Cell Cas12a Platform for Screening and CRISPRi

To overcome the challenges of pre-crRNA sequence recovery during single-cell RNA sequencing, we optimized guide RNA expression vectors and established a degron-based, enhanced Cas12a system for scalable genetic perturbation. Using this new platform, we performed Perturb-seq screens to accurately capture pre-crRNAs and profile gene editing-induced effects on the transcriptome across multiple cell types and target genes. This dataset provides the single-cell transcriptomic profiles demonstrating the efficacy of our improved Cas12a and HyperLbCas12a systems for highly multiplexed gene knock-out and transient suppression.