Degron-modified Cas12a enhances single-cell CRISPR screening

Sep 1, 2024·
Valentina Snetkova
,
Carolina Galan
,
Romain Lopez
,
Antonio R. Rios
,
Takamasa Kudo
,
Kristel Dorighi
,
Søren Warming
,
Benjamin J. Haley
· 0 min read
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 Cas12a’s 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.
Type
Publication
bioRxiv