产品说明
一般描述
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).
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
- 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.
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
All-in-One is a trademark of sigma-aldrich Co. LLC
基本信息
| NACRES | NA.51 |
产品性质
| 质量水平 | 200 |
| 包装 | pkg of 8x25 μL (vials) |
| 浓度 | 5x108 VP/ml (via p24 assay) |
| application(s) | CRISPR |
| 运输 | dry ice |
| 储存温度 | −70℃ |
安全信息
| 储存分类代码 | 12 - Non Combustible Liquids |
| WGK | WGK 3 |
| 闪点(F) | Not applicable |
| 闪点(C) | Not applicable |
