Perform unbiased, phenotypic knockout screens using Edit-R Lentiviral sgRNA Pooled Screening Libraries without the need for costly infrastructure.
Loss-of-function screens are powerful forward genetic approaches for discovering novel gene functions and interactions in both normal and disease biology. The recent exciting discoveries and application of CRISPR-Cas9 gene editing mechanisms in eukaryotic cells has facilitated the ability to knockout hundreds and even thousands of genes at the DNA level through pooled lentiviral library screening methods.
The ability of any given CRISPR guide RNA to create a double-strand break in target DNA that causes functional protein disruption can vary based on the guide RNA (gRNA) sequence and position in the targeted gene. Likewise, the overall specificity of RNA-directed DNA cleavage events is not yet completely understood and can hamper its wider application. To address this, Dharmacon has developed a proprietary algorithm for genome-wide design of highly specific and functional RNA guides across the human, mouse and rat genomes. The Edit-R CRISPR RNA algorithm generates guide RNA sequences with:
unparalleled specificity through an optimized alignment program which performs rapid and thorough specificity analysis, including detection of multiple mismatches and gapped alignments
increased functional knockout of targeted genes through systematic assessment of over 1100 guide RNAs to identify characteristics critical for functional gene disruption, not just generation of indels.
Using the Edit-R CRISPR RNA algorithm, we have designed lentiviral sgRNAs showing highly efficient gene knockout at single-copy integration levels required for pooled lentiviral screens.
Efficient two-vector system that utilizes a lentiviral vector for Cas9 expression and a gene-specific vector for sgRNA expression
Rationally designed Edit-R lentiviral sgRNAs efficient gene knockout with unparalleled specificity using a functionally validated proprietary algorithm
Deep and broad coverage of 5 -10 sgRNAs per gene across the human, mouse and rat genomes for increased hit confidence and comprehensive genome screening
Can be combined with the promoter flexibility of Edit-R Lentiviral Cas9 Nuclease Reagents for robust editing in biologically relevant cell types
Each Edit-R Lentiviral sgRNA Pooled Screening Library includes
- ≥ 5 x 108 TU/mL ( ± 20%) lentiviral particles in pre-aliquoted tubes
- 8 x 25 µL (200 µL total) for libraries ≤ 5000 constructs
- 16 x 25 µL (400 µL total) for libraries > 5000 constructs
- Five to ten sgRNAs per gene targeting coding genes in the NCBI Reference Sequence Database
- 100 non-targeting sgRNA negative controls bioinformatically confirmed to not align with (target) any gene in the human, mouse and rat genomes.
- Up to 340 gene-specific sgRNA positive controls targeting up to 34 protein-coding genes (10 sgRNA per gene); including common reference genes (ACTB, GAPDH, LAMB1, & PPIB)
- A data file containing complete library information, including: gene annotations, sgRNA target sequences, complete list of controls, and counts per millions of mapped reads
Edit-R Lentiviral sgRNA libraries are available for defined sub-libraries of gene families up to the entire genome for human and mouse; rat libraries and custom Edit-R Lentiviral sgRNA collections are available upon request.
- Request a custom collection of human, mouse, or rat gene-targeting sgRNAs
- Available in pool sizes between 50 to 10,000 constructs
- Pools are provided as purified, concentrated lentiviral particles ≥ 5 x 108 TU/mL
Products recommended in our validated protocol
- Vector-matched Edit-R Lentiviral sgRNA Non-targeting Control for transduction optimization
- Edit-R Pooled sgRNA Indexing PCR and Sequencing Primer Kits (A & B) with 12 unique indexing primers each, optimized and experimentally validated for:
- Efficient PCR amplification of genomic DNA with minimal bias
- High-throughput multiplexed Next Gen Sequencing on an Illumina flow cell for hit identification
Edit-R Lentiviral sgRNA pooled library coverage*
* The number of genes and constructs per pool are subject to change at any time without notice.
** Clone and gene counts available by request
Edit-R Lentiviral sgRNA Pooled Library||
# targeted genes||
Number of pools and average constructs per pool||
# targeted genes||
Number of pools and average constructs per pool|
18 pools of 10,450 constructs||
20 pools of 10,300 constructs|
7 pools of 10,730 constructs||
10 pools of 10,170 constructs|
1 pool of 4127 constructs||
1 pool of 5311 constructs|
1 pool of 3829 constructs||
1 pool of 3818 constructs|
1 pool of 7262 constructs||
1 pool of 7451 constructs|
1 pool of 2846 constructs||
1 pool of 3089 constructs|
1 pool of 5080 constructs||
1 pool of 5712 constructs|
1 pool of 5642 constructs||
1 pool of 5564 constructs|
Cell Cycle Regulation||
DNA Damage Response||
The Edit-R Lentiviral sgRNA Pooled Libraries are solely for internal research use (as set forth in the Product Terms and Conditions) in laboratories where the containment measures stated below and in applicable laws and regulations are met. Products may not be used for diagnostic, therapeutic or other commercial purposes and may not to be administered to humans for any purpose or to animals for therapeutic purposes. Edit-R Lentiviral sgRNA Pooled Libraries are provided as lentiviral particles are replication-incompetent, self-inactivating (SIN) and non-pathogenic (do not cause infectious human disease).
Any investigator who purchases lentiviral particle products is responsible for consulting with their institution's health and biosafety personnel for specific guidelines on the handling of lentiviral vector particles. Furthermore, each investigator is fully responsible for obtaining the required permissions for research using and the acceptance of replication-incompetent SIN lentiviral vectors and replication-defective lentiviral particles into their local jurisdiction and institution.