Scalable single-cell pooled CRISPR screens with conventional knockout vector libraries
Islam, M.; Yang, Y.; Simmons, A. J.; Xu, Y.; Fisher, E.; Deng, W.; Grieb, B.; Molina, P.; Caestecker, C. d.; Ramirez-Solano, M. A.; Liu, Q.; Tansey, W.; Macara, I. G.; Rathmell, J.; Coffey, R. J.; Lau, K.
Show abstract
Current methods for single-cell RNA profiling of pooled CRISPR screens are limited, either by indirect capture of single guide RNAs (sgRNAs) or by custom modification of plasmid libraries. Here, we present a direct sgRNA capture platform called Native sgRNA Capture and sequencing (NSC-seq) that enables single-cell CRISPR screens using common knockout plasmid libraries, facilitating genotype-phenotype mapping at multiple scales in vitro and in vivo. Additionally, we characterize sgRNA expression in three whole-genome knockout libraries, revealing a substantial subset of truncated (isoform) spacer reads. We provide this dataset as a reference of expressed sgRNA isoforms that may potentially have compromised CRISPR gene editing efficacy and precision.
Matching journals
The top 5 journals account for 50% of the predicted probability mass.
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.