Degron-modified Cas12a enhances single-cell CRISPR screening
Snetkova, V.; Galan, C.; Lopez, R.; Rios, A. R.; Kudo, T.; Dorighi, K.; Warming, S.; Haley, B. J.
Show abstract
Single-cell CRISPR (Perturb-seq) screens have primarily relied on Cas9 whereas Cas12a, despite its unique effectiveness for multiplex guide expression, remains underexplored. This may be due to Cas12as guide RNA array (pre-crRNA) self-processing activity and the subsequent challenges associated with pre-crRNA sequence recovery. By developing modified pre-crRNA expression vectors and a degron-based Cas12a system, we overcome the self-processing constraint, allowing for accurate detection of pre-crRNAs at the single-cell level, thus greatly expanding possibilities for future Perturb-seq efforts.
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