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MGECKO2G

Sigma-Aldrich

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

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About This Item

Code UNSPSC :
12352200
Nomenclature NACRES :
NA.51

Conditionnement

pkg of 8x25 μL (vials)

Niveau de qualité

200

Concentration

5x108  VP/ml (via p24 assay)

Application(s)

CRISPR

Conditions d'expédition

dry ice

Température de stockage

−70°C

Catégories apparentées

Description générale

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).

Application

Functional Genomics/Screening /Target Validation

Caractéristiques et avantages

  • 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.

Notes préparatoires

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.

Autres remarques

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.

Informations légales

CRISPR Use License Agreement

Lentiviral License Agreement

Code de la classe de stockage

12 - Non Combustible Liquids

Classe de danger pour l'eau (WGK)

WGK 3

Point d'éclair (°F)

Not applicable

Point d'éclair (°C)

Not applicable

Certificats d'analyse (COA)

Recherchez un Certificats d'analyse (COA) en saisissant le numéro de lot du produit. Les numéros de lot figurent sur l'étiquette du produit après les mots "Lot" ou "Batch".

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