Optimizing an Arrayed CRISPR Screen Arrayed CRISPR screens are powerful functional genomics tools that can interrogate thousands of genes in parallel. Optimizing aspects of your screening workflow is a critical first step. In this eBook, we provide step-by-step instructions for optimizing the delivery...
(a–c) Pairwise comparisons of significant (FDR < 0.01) and not significant results among the three replicates (Rep) of Daudi-Cas9 cell populations used for the BTN3A expression screen. (d) Correlation of screen effect sizes (LFC) among concordant hits separated into positive and negative regu...
A surfaceome CRISPR activation screen for candidate genes that confer resistance to NK cell cytotoxicity We conducted a surfaceome-focused CRISPRa screen of K562 cells to identify tumor ligands that modulate NK cell-mediated killing. K562 cells are a NK-sensitive chronic myelogenous leukemia cell lin...
8a–c). We then exposed these models to the drugs (for the FFA-induced steatosis model, still in the presence of FFAs) for 7 days. Lipid accumulation was assessed through Nile Red staining, and we developed a lipid droplet scoring system to score drug effectiveness (Fig. 3a and ...
Missense/nonsense mutations are widely spread over in tumor-associated genes [147,148]. Harnessing the unique base conversion capability of BEs can reprogram missense mutations and nonsense mutations of ORFs [149,150]. Therefore, BEs can be used as a strategy to accurately screen [151,152,153,...
(LDL-C) without causing any clinical risk. Qiu et al. designed multiple LNPs for the delivery of Cas9 mRNA and an sgRNA targeting Angptl3.195,196A gold standard MC-3 LNP configured with cholesterol, DSPC and DMG-PEG was used as a control to screen for the most efficient 306-O12B LNP ...
We transduced both libraries, in duplicate, at a depth-coverage of 20,000 (KPT-9274) or 10,000 (ispinesib) cells per sgRNA into HAP1 cells stably expressing SpCas9-NG to screen for drug resistance against KPT-9274 and ispinesib (Figure S2A). Our original results with SpCas9 indicated...
Cloning of orthologues for activity screen and recombinant expression We synthesized human codon-optimized versions of 15CRIPSRCas13a orthologues (Genscript) and cloned them into pACYC184 under a pLac promoter. Adjacent to the Cas13a expression cassette, we cloned the orthologue’s corresponding direct...
An arrayed CRISPR activation screen for TFs controlling PrPC expression The cellular prion protein PrPC, encoded by the PRNP gene, is required for the development of prion diseases37,38. Previous genome-wide microRNA and siRNA screens have uncovered a complex pattern of regulated expression of PrPC...
A surfaceome CRISPR activation screen for candidate genes that confer resistance to NK cell cytotoxicity We conducted a surfaceome-focused CRISPRa screen of K562 cells to identify tumor ligands that modulate NK cell-mediated killing. K562 cells are a NK-sensitive chronic myelogenous leukemia cell lin...