CRISPR/Cas9 Genome-Wide gRNA Library for Target Identification
6 Jun 2017Genome-wide loss-of-function screening is a fundamental method to identify genes responsible for driving biological responses, and complex pooled lentiviral-based libraries expressing large numbers of genetic disruptors, such as shRNAs, make large-scale cell screening practical. This study investigates the potential differences in the RNAi and CRISPR HTGS technologies, by performing parallel dropout viability screens using module-1 human genome-wide CRISPR sgRNA library and module-1 human genome-wide RNAi shRNA library, both modules targeting the same set of 6,300 genes with the same number of effectors (sgRNA or shRNA) for each gene.
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CRISPRtest™ Functional Cas9 Activity Kit
CellectaWant to know if Cas9 is active in your cells? Cellecta’s CRISPRtest™ method allows you to assess the level of Cas9 activity in your cell line using two pre-mixed lentiviral-packaged vectors, the first containing a green fluorescent protein (GFP) marker and gRNA sequence targeting an essential gene (PCNA), and the second containing a red fluorescent protein (RFP) marker and a non-targeting gRNA. Transduction of CRISPRtest virus into cells will result in a mixture of cells expressing GFP and cells expressing RFP. Upon cell growth in the presence of active Cas9, the ratio of GFP cells: RFP cells will decrease proportionally to Cas9 activity. The CRISPRtest Functional Cas9 Activity Kit, currently for human cell lines only, is an easy-to-use, highly sensitive kit for measuring functional Cas9 activity in almost any human cell line.
CRISPR Human Genome 80K Knockout Library (plasmid)
CellectaCellecta’s 80K Single-module Human Genome-Wide CRISPR Library was designed to be small enough (~80,000 constructs) to enable convenient genome-wide “dropout viability” screens with most standard human cell modules. With 4 sgRNAs targeting each gene, all of which have been designed using the latest criteria for effective guides described by Doench, et al. and containing the Cellecta-designed HEAT sgRNA structure which we have proven to generate more robust screening results than the standard sgRNA design, the library has deep enough gene coverage to generate efficient results. Genome-Wide Coverage with a Convenient Single 80K Module Single library with ~80,000 constructs targeting approximately 19,000 protein-coding genes Each gene is targeted by 4 sgRNAs designed based on Doench, et al. criteria Targets all human protein-coding genes—over 19,000 gene targets. The 80K Single-Module Human Genome-Wide CRISPR Library includes the following controls standard non-targeting controls non-specific control (sgRNA targeting introns throughout the genome) lethal sgRNA targeting generally essential genes (i.e., positive controls for dropout screens)