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HWGCRISPR

Sigma-Aldrich

Sigma Whole Human Genome Lentiviral CRISPR Pool

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About This Item

UNSPSC Code:
41106609
NACRES:
NA.51

packaging

pkg of 8x25 μL (vials)

Quality Level

200

concentration

5x108  VP/ml (via p24 assay)

application(s)

CRISPR

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.

Legal Information

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Campylobacter Ecology and Evolution (2014)

Articles

Safe Lentivirus Handling

Our lentiviral vector systems are developed with enhanced safety features. Numerous precautions are in place in the design of our lentiviruses to prevent replication. Good handling practices are a must.

Successful Transduction Using Lentivirus

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.

Protocols

CRISPR Lentiviral Screening: Antibiotic Selection

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.

FACS Titration of Lentivirus Protocol

FACS (Fluorescence-Activated Cell Sorting) provides a method for sorting a mixed population of cells into two or more groups, one cell at a time, based on the specific light scattering and fluorescence of each cell. This method provides fast, objective, and quantitative recording of fluorescent signals from individual cells.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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