MGECKO2G

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

• 分類程式碼代碼: 12352200
• NACRES: NA.51

属性

• 包裝: pkg of 8x25 μL (vials)
• 品質等級: 200
• 濃度: 5x10⁸ VP/ml (via p24 assay)
• 應用: CRISPR
• 運輸包裝: dry ice
• 儲存溫度: −70°C

说明

一般說明

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).

應用

Functional Genomics/Screening /Target Validation

特點和優勢

• 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.

準備報告

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.

其他說明

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。

法律資訊

CRISPR专利正在申请中

安全信息

• 儲存類別代碼: 12 - Non Combustible Liquids
• 水污染物質分類（WGK）: WGK 3
• 閃點（°F）: Not applicable
• 閃點（°C）: Not applicable