For combinatorial screens, a screen of previously reported synthetic lethal paralog pairs suggested a design of just six constructs per gene pair was sufficient to identify these genetic interactions.

For single-target screens, just one construct, the top a priori ranked guide pair from Broad GPP’s CRISPick, provides strong performance.

We observed robust performance for both guide positions, all three validated tRNAs, and with diverse 12 nt iBAR sequences.

Second, we validate a scRNA-seq readout based on 5’ direct-capture with high capture rates.

Our updated design, CROPseq-multi-v2 (Addgene: www.addgene.org/225752/), retains all the functionality of the original CROPseq-multi, while adding compatibility for methods based on T7-IVT-based barcode detection approaches, like PerturbView and NIS-Seq for optical pooled screens.

We’re sharing a major update to CROPseq-multi, our versatile system for CRISPR screens that is compatible with individual and combinatorial perturbations, diverse SpCas9-based technologies, and multiple high-content, single-cell readouts. www.biorxiv.org/content/10.1...