CRISPR Human Genome-Wide Knockout Libraries
Quality-checked pooled lentiviral libraries with mutation rates less than 0.5% and at least 90% of the sgRNA represented within a 5-fold distribution range. Both plasmid and pre-packaged CRISPR Human Genome-Wide Knockout Libraries are available.
- Effective guides incorporate Cellecta’s tracrRNA “HEAT” design and have been screened to minimize off-target knockout
- Both plasmid and pre-packaged lentivirus forms of the libraries are available
- Sufficient plasmid library to use immediately for packaging (no reamplification necessary)
- Two-vector CRISPR system with a strongly expressing Cas9 plasmid as one component, and a library of sgRNA-expressing constructs as the second.
Cellecta, the first company to release a genome-wide lentiviral based CRISPR library, continues to offer quality pooled lentiviral CRISPR libraries for unbiased phenotypic knockout screens in mammalian cell systems. Pooled libraries of many thousands of defined single-guide RNA (sgRNA or gRNA) sequences enable disruption (or “knockout”) of thousands of specific genes in a single experiment. This loss-of-function genetic screen platform enables, for example, the identification of specific genes required for a biological response as well as of potential drug targets for disease phenotypes.
In keeping with our mission to provide accessible advanced technology to accelerate functional genomic analysis, Cellecta provides CRISPR libraries in both plasmid and pre-packaged forms with large portions for immediate use. All libraries are provided with complete sequence information on the vectors, inserts, and guides expressed, as well as Next-Generation Sequencing (NGS) data showing the number of reads per sgRNA as well as the distribution of the sgRNA of the library.
Guide designs for Cellecta’s libraries incorporate state-of-the-art design criteria to ensure the most effective knockout possible. In addition, the region of the sgRNA interacting with the Cas9 nuclease has been optimized for higher efficiency knockout by incorporating the “HEAT” design. We have shown that specific modifications to the Cas9-binding region of sgRNA significantly improve the effectiveness of gene knockout and that CRISPR libraries containing constructs with these modified sgRNA sequences generate stronger and more robust results than those with the standard sgRNA.
Cellecta offers two pre-made CRISPR Human Genome-wide Knockout Libraries. Both library options cover all human protein-coding genes:
- 80K Human Genome-Wide CRISPR Knockout Library
Genome-wide coverage in a single, pooled library of approximately 80,000 constructs that can be conveniently used for a wide variety of screening approaches.
- 3-Module Human Genome-Wide Knockout Library
Deep coverage of 8 sgRNA per target gene divided into 3 modules that may be combined or used separately, depending on the type of screen desired.
These off-the-shelf libraries are available for convenient ordering online.
Figure 1. sgRNA Structure Optimization Screen Data: Waterfall plots display the depletion fold of positive and negative controls within the library for each design. The HEAT design shows best performance.
Figure 2. Z-scores show improvements achieved with HEAT sgRNA handle design, used for Cellecta’s CRISPR Human Genome-wide Knockout Libraries.