一般描述
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).
应用
Functional Genomics/Screening /Target Validation
特点和优势
- 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.
制备说明
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.
其他说明
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。
法律信息
CRISPR专利正在申请中
WGK
WGK 3
闪点(°F)
Not applicable
闪点(°C)
Not applicable
我们的科学家团队拥有各种研究领域经验，包括生命科学、材料科学、化学合成、色谱、分析及许多其他领域.
联系技术服务部门