• USA Home
  • HWGCRISPR - Sigma Whole Human Genome Lentiviral CRISPR Pool

EMAIL THIS PAGE TO A FRIEND
HWGCRISPR Sigma-Aldrich

Sigma Whole Human Genome Lentiviral CRISPR Pool

Purchase

Properties

Related Categories CRISPR-Cas9, Functional Genomics and RNAi, Lentiviral CRISPR Pools, Molecular Biology
application(s)   CRISPR: suitable
shipped in   dry ice
storage temp.   −70°C

Description

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.

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.

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

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.

Legal Information

CRISPR Use License Agreement

Lentiviral and WPRE License Agreements

Safety & Documentation

Safety Information

RIDADR 
UN 3245 9

Documents

Certificate of Analysis


Making Headway
Protocols & Articles

Articles

Safe Handling of Lentivirus

Our lentiviral vector systems have been developed with enhanced safety features. Numerous precautions are in place in the design of our lentiviruses to prevent replication, including eliminating viru...
Keywords: Environmental, Genetic, Recombination, Transfection

Protocols

CRISPR Lentiviral Screening: Antibiotic Selection

A kill curve is a dose-response experiment where cells are subjected to a range of antibiotic concentrations to determine the minimum concentration of antibiotic needed to kill all cells over a desir...
Keywords: Antibiotics, Apoptosis, Gene expression, Transduction

Protocol, FACS Titration of Lentivirus

FACS Analysis of CFU 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 scat...
Keywords: Enzyme-linked immunosorbent assay, Flow cytometry, Gene expression, Titrations, Transduction

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. As...
Keywords: Antibiotics, Enzyme-linked immunosorbent assay, Gene expression, PAGE, Polymerase chain reaction, Titrations, Transduction

Related Content

CRISPR

In recent years CRISPR has revolutionized gene editing capabilities, leading to sophisticated ways to create success with any experiment. As the first company to offer custom biomolecules globally fo...
Keywords: Genetic, Transduction, Transfection

Predictive Models for Neuroscience using CRISPR [VIDEO]

Caroline Beckett, the global CRISPR product manager for MilliporeSigma, discusses reagent solutions for creating predictive models for neuroscience research. She notes that many neuroscientists want ...
Keywords: Cell culture, Diseases, Neurodegenerative Diseases, Neuroscience

Peer-Reviewed Papers
15

References

Related Products

Technical Service:

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

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?