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

半岛bd体育手机客户端 说明

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).

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

该半岛bd体育手机客户端 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

半岛bd体育手机客户端 性质

质量水平	200
包装	pkg of 8x25 μL (vials)
浓度	5x108VP/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