HWGCRISPR

Sigma-Aldrich

Sigma Whole Human Genome Lentiviral CRISPR Pool

NACRES:
NA.51

Quality Level

packaging

pkg of 8x25 μL (vials)

concentration

5x108  VP/ml (via p24 assay)

application(s)

CRISPR: suitable

shipped in

dry ice

storage temp.

−70°C

General description

CRISPR (clustered regularly interspaced short palindromic repeat) are repeated sequence of DNA. These are transcribed in a single transcript and later divided into short repeats.

Application

Functional Genomics/Screening /Target Validation

Biochem/physiol Actions

Utilize the power of CRISPR Whole Genome Lentiviral Pools to knockout and screen every gene in the human genome. The Sigma Whole Human Genome Lentiviral CRISPR Pool is provided as 200 ul in 8 x 25 ul aliquots at a minimum titer of 5x108 viral particles/ml via p24.

Features and Benefits

  • Use CRISPR nucleases to knockout protein-coding genes to assess their function
  • Efficiently screen the whole human genome (16,000+ genes) at the bench-top without robotics or specialized equipment
  • Increased flexibility consisting of 8 subpools at 23,000 clones per pool (184,000 clones total), from the previous 2 subpools (Gecko v2)
  • Expanded gRNA coverage at 10 gRNAs per gene, from the previous 6 gRNAs per gene (Gecko v2)
  • Numerous built-in enrichment and depletion controls allows researchers to confidently gauge the success of their pooled screening experiments
  • 2 Vector System (pools are gRNA-only, Cas9 sold separately)
  • Ease of optimization: Utilizes same vector system as Gecko v2 allowing for optimization one for both systems
  • Minimize off-targeting: stringent gRNA design and tiling rules

Preparation Note

Puro Kill Curve and Determining CFU (Colony Formation Unit) per mL.
Prior to performing a library-scale screening, two preliminary experiments must be conducted: (1) determine the sensitivity of target cell type to puromycin (kill curve), and (2) determine the functional titer of the lentivirus in your cell type by completing a colony-forming assay (measured in CFU/ml). The calculation of MOI (multiplicity of infection) should be based on the value of CFU. Different cell types vary in transduction efficiency and different lentiviral constructs do not behave identically, so it is critical to optimize your experimental conditions with control lentiviral CRISPR clones (available from Sigma) prior to performing your pooled experiment.

Other Notes

This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. Though the lentiviral transduction particles produced are replication incompetent, it is recommended that they be treated as Risk Group Level 2 (RGL-2) organisms in laboratory handling. Follow all published RGL-2 guidelines for laboratory handling and waste decontamination.

storage_class_code

12 - Non Combustible Liquids

WGK Germany

WGK 3

Flash Point F

Not applicable

Flash Point C

Not applicable

Certificate of Analysis

Certificate of Origin

Campylobacter Ecology and Evolution (2014)
Articles
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Successful targeting relies on optimizing key sensitive steps in the process, including lentiviral transduction. Below are some helpful handling and titration tips from our R&D lentiviral experts.
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Protocols
You are not alone designing successful CRISPR, RNAi, and ORF experiments. Sigma-Aldrich was the first company to commercially offer lentivirus versions of targeted genome modification technologies and has the expertise and commitment to support new generations of scientists.
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