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- 文献和实验
- 技术资料
- 库存：
大量
- 英文名：
pLV-EF1-dCas9-VPR-PGK-PURO
- 保质期：
详询
- 供应商：
北京百奥创新科技有限公司
- 保存条件：
低温
- 规格：
10 μg at 0.5 μg/μL in TE
Description:CRISPR (clustered regularly interspaced short palindromic repeats) activation (CRISPRa) is a widely used genetic technique that allows for targeted activation of transcription in bacteria and mammalian cells. The CRISPRa system is comprised of a catalytically inactive Cas9 (dCas9) protein fused with a transcriptional activator such as the tripartite fusion of three transcription activation domains: VP64, p65 and Rta (VPR) and a customizable single guide RNA (sgRNA). The Cas9-sgRNA complex binds to DNA elements complementary to the sgRNA and recruits transcription factors to increase gene expression, resulting in the activation of the target gene.
pLV-EF1-dCas9-VPR-PGK-PURO Lentiviral Vector expresses the dCas9-VPR fusion with an EF1 promoter and PURO with a PGK promoter to allow for selection of transduced cells.
Vector Information：This is a lentiviral expression vector that contains all the elements necessary for efficient and high yield viral production. Ubiquitous EF1 and PGK promoters drive the expression of the dCas9-VPR fusion protein and PURO, respectively, to allow for the selection of transduced cells. This vector can be used for stable cell line generation or in combination with a gene specifc sgRNA to activate the target gene expression.
IMPORTANT NOTICE：Store the vial at -20°C immediately upon receipt.
Specifications:
| Product Name | pLV-EF1-dCas9-VPR-PGK-PURO |
|---|---|
| Shipping Condition | Room Temperature - 2 Day Shipping |
| Storage and Stability | Store at -20°C immediately upon receipt. This product is stable for 6 months when stored as directed. |
| Quality Control | This plasmid is sequence verified. |
| Restricted Use | For Research Use Only. Not for use in diagnostic or therapeutic procedures. |
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文献和实验会怎样"用 CRISPRa。 --- 一、机制差异决定了两者能回答的问题不同 过表达质粒（cDNA / ORF） 外源启动子（CMV、EF1α 等）驱动，表达量与内源调控完全脱钩； 通常只包含单一 CDS，不含内含子和 UTR； 表达量可达到内源的几十到几百倍； 可携带标签（FLAG、HA、GFP）方便检测。 CRISPRa（dCas9-VP64 / VPR / SAM / SunTag） 失活 Cas9（dCas9）融合转录激活域，被 gRNA 引导至靶基因启动子区； 激活的是内源位点，转录本包含完整的内含
一、试剂准备 1、Cas9 过表达慢病毒载体选择。 货号 名称 CR2001 pLV-Cas9-Puro CR2002 pLV-Cas9-Neo CR2003 pLV-Cas9Nick-Puro CR2004
-of-function）是让基因表达变多或变强，常用三招：直接构建过表达载体、CRISPR-SAM 激活（CRISPRa）、saRNA 上调。功能缺失（loss-of-function）是让基因表达变少或彻底没，常用：RNAi 干扰（shRNA/siRNA）、CRISPR-Cas9 敲除、cas13d 干扰、dCas9-KRAB 转录抑制。优先级一般是 A>B>C>D，以你的实际需求为导向来选，不是越新越好。说白了，一类是"加量"，一类是"减量或撤掉"，方向完全相反。 它们分别在细胞里怎么跑 过表达是把编码序列连到
