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MGECKO2A Sigma

Gecko2 Mouse Whole Genome CRISPR Pool, All-in-one Lenti Particles (Gecko2 vector)

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

Description

General description

Developed by Feng Zhang′s lab at the Broad Institute, the mouse GeCKO v2 (all-in-one vector) libraries consist of over 100,000 unique gRNAs for gene knock-out in the mouse genome. Using an all-in-one lenti CRISPR vector which co-expresses Cas9 , gRNA with a puromycin selection marker, the version 2 vector produces nearly 10X higher titer than version 1. Sigma′s lentiviral mouse GeCKO pool all-in-one is provided in 8 x 25 ul aliquots at a minimum titer of 5X10^8 TU/ml (measured by a p24 assay).

Each species-specific library is delivered as two half-libraries (A and B). It is recommended to screen both A and B libraries together, which will include 6 sgRNAs per gene (3 sgRNAs in each library). Both libraries contain 1000 non-targeting control sgRNAs. The A library also targets miRNAs (4 sgRNAs per miRNA).

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. Visit Sigma.com/pooledscreening.

Legal Information

CRISPR Use License Agreement

Lentiviral License Agreement

Application

Functional Genomics/Screening /Target Validation

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

Numerous built-in enrichment and depletion controls allow researchers to confidently gauge the success of their pooled screening experiments
• Use the one vector system for the mouse GeCKO version 2 libraries in applications where the Cas9 cannot be introduced into mouse cells (like in primary cells).
• Lentiviral CRISPRs can infect a broad variety of mammalian cells by co-expressing a mammalian codon-optimized Cas9 nuclease along with a single guide RNA (sgRNA) to facilitate gene knockout.
• Use puromycin to select after transduction.

Safety & Documentation

Safety Information

RIDADR 
UN3373 - class 6.2 Biological substance, Category B
WGK Germany 
3

Documents

Certificate of Analysis

Protocols & Articles

Protocols

Protocol Guide: CRISPR/CAS9 Gene Editing of Human Induced Pluripotent Stem Cells (iPSCs)

Induced pluripotent stem cells (iPSCs), have the capacity to give rise to differentiated progeny arising from of all germ layers of the body including: ectoderm, endoderm, and mesoderm. The ability t...
Keywords: Alzheimer Disease, Apoptosis, Cell culture, Centrifugation, Cloning, Culture media, Gene expression, Genetic, Microscopy, Parkinson Disease, Phase transitions, Transfection

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Predictive Models for Neuroscience using CRISPR [VIDEO]

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