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MGECKO2G Sigma

Gecko2 Mouse Whole Genome CRISPR Pool, gRNA Only Lenti Particles (Gecko2 vector)

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Properties

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 (two vector system) libraries consist of over 100,000 unique gRNAs for gene knock-out in the mouse genome. Using a dual lenti CRISPR vector wherein the pooled libraries will only express the gRNA (with the lentiGuide-Puro vector), this system produces over 100X higher titer virus compared to version 1. The lentiGuide-Puro pool should be used only in cell lines with Cas9 already integrated (which can be generated using a separate lenti-Cas9-Blast vector). Sigma′s lentiviral mouse GeCKO pool guide RNA only 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 • Lentiviral CRISPRs can infect a broad variety of mammalian cells by transducing a single guide RNA (sgRNA) to a Cas9-expressing mouse cell line to facilitate gene knockout for screening applications.
• Use the dual vector system for the mouse GeCKO version 2 libraries for mouse cell lines that have Cas9 already integrated into the genome.
• Use puromycin gRNA selection after transduction.

Safety & Documentation

Safety Information

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

Documents

Certificate of Analysis

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

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 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

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

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

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