Perform unbiased, phenotypic knockout screens without the need for costly infrastructure

Knockout hundreds and even thousands of genes at the DNA level through pooled lentiviral library screening methods.

The ability of CRISPR guide RNA to create a double-strand break in target DNA causing functional protein disruption can vary based on the guide RNA (gRNA) sequence and position in the targeted gene. To address this, the Dharmacon reagents algorithm provides highly specific and functional RNA guides across the human genome and mouse genomes.

The Edit-R CRISPR RNA algorithm generates guide RNA sequences with:

• An optimized alignment program with 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
• New! Edit-R human sgRNA designs have been updated to the latest RefSeq in 2025 providing the most specific and genomically relevant guides for producing efficient protein knockout. This allows the Edit-R algorithm to target the latest genome annotations more accurately and efficiently providing you with the best solution for your research needs. Please reach out to Scientific Support if you have any questions.

Highlights:

• Efficient two-vector system utilizing 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 4 or 8 sgRNAs per gene across the human genome for whole genome library or 5 -10 sgRNAs per gene across the mouse genome for increased hit confidence.
• Can be combined with the promoter flexibility of Edit-R Lentiviral Cas9 Nuclease Reagents for robust editing in biologically relevant cell types

The NEW Edit-R Human Lentiviral sgRNA Pooled Screening Libraries include, per pool:

• ≥ 5 x 10⁸ TU/mL ( ± 20%) lentiviral particles in 10 x 100 uL pack size
• Four or eight sgRNAs per gene (whole genome) and up to four sgRNAs per gene (gene family and pathway libraries) 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 genome
• Gene family and pathway libraries include up to 136 gene-specific sgRNA technical controls targeting up to 34 protein-coding genes (4 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

The Edit-R Mouse Lentiviral sgRNA Pooled Screening Libraries include, per pool:

• ≥ 5 x 10⁸ TU/mL ( ± 20%) lentiviral particles in pre-aliquoted tubes

• 8 x 25 µL (200 µL total) for pools ≤ 5000 constructs
• 16 x 25 µL (400 µL total) for pools > 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 and mouse 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

Products recommended in our validated protocol:

• Vector-matched Edit-R lentiviral positive and non-targeting sgRNA controls for gene editing validation and experimental optimization
• SMARTvector Non-targeting Control Particles with optimal promoter and fluorescent reporter (TurboGFP or TurboRFP) for determining relative transduction efficiency and optimizing transduction conditions in desired cells of interest.
• Edit-R Lentiviral Cas9 Nuclease Reagents or Cas9 Stable Cell Line

• Human libraries in NEW V2 backbone:
  • NEW: NGS Library Prep Kit (Recommended)
  • Primer set options available two different sizes for 12 or 24 whole genome screens
  • Designed to amplify and sequence sgRNA constructs from gDNA without bias
  • One complete solution for sequencing on an Illumina® platform without the need for additional reagents
• Mouse libraries and Human libraries in V1 backbone: 
  • Edit-R Mouse Pooled sgRNA Indexing PCR and Sequencing Primer Kit A for  Illumina® sequencing platforms (required)
  • Edit-R Mouse Pooled sgRNA Indexing PCR and Sequencing Primer Kit B for additional reverse indexing PCR primers (if experimental design requires additional multiplexing capability)

Important Notice

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 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.

** Please contact Scientific Support for more information