packaging
pkg of 8x25 μL (vials)
concentration
5x108 VP/ml (via p24 assay)
application(s)
CRISPR
shipped in
dry ice
storage temp.
−70°C
Quality Level
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.
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).
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).
Legal Information
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.
存储类别
12 - Non Combustible Liquids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
法规信息
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Lentiviral vector systems prioritize safety features, with design precautions preventing replication. Good handling practices are essential for use.
Get tips for handling lentiviruses, optimizing experiment setup, titering lentivirus particles, and selecting helpful products for transduction.
获取处理慢病毒、优化实验设置、测定慢病毒颗粒滴度以及选择实用转导产品方面的贴士。
实验方案
Lentivirus versions of genome modification technologies support successful CRISPR, RNAi, and ORF experiments.
FACS sorts cells based on light scattering and fluorescence for objective cell analysis.
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