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bioRxiv · 10.1101/2024.12.08.627374

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.
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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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Snetkova, V., Galan, C., Lopez, R., Rios, A. R., Kudo, T., Dorighi, K., Warming, S., Haley, B. J.. 2024-12-08. Degron-modified Cas12a enhances single-cell CRISPR screening. https://doi.org/10.1101/2024.12.08.627374

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