Gecko2 Mouse Whole Genome CRISPR Pool, All-in-One^™ Lenti Particles (Gecko2 vector)

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).

Functional Genomics/Screening /Target Validation

• 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.

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.

该产品仅供研发使用，不可用作药物、家庭使用或其他用途。请查阅物质安全资料表，获取危险和安全操作规范信息。尽管生产的慢病毒转导颗粒是不可复制的，但仍然建议将其作为生物危害等级2级（RGL−2）生物体在实验室中进行处理。请遵循所有已发表的RGL-2指南，进行实验室废物处理和净化。

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