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- 英文名:
pLVX-shRNA1
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- 供应商:
武汉华尔纳生物科技有限公司
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Product Introduction
| 品牌 |
华尔纳生物 | ||||||||||||||||||||||||
| 货号 |
WN-12839 | ||||||||||||||||||||||||
| 规格 |
2ug | ||||||||||||||||||||||||
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询价 | ||||||||||||||||||||||||
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现货 | ||||||||||||||||||||||||
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质粒图谱 载体描述描述pLVX-shRNA1 is an HIV-1-based, lentiviral expression vector designed to express a small hair-pin RNA (shRNA) for RNA interference (RNAi) studies. Expression of your shRNA is drivenby the RNA Pol III-dependent, human U6 promoter (PU6), located just upstream of the MCS. pLVX-shRNA1 can either be used as a plasmid expression vector and transfected into cells, or it can be packaged into viral particles and transduced into cells. Lentiviral particles derived from the vector allow the expression of shRNAs in virtually any cell type, including primary cells. pLVX-shRNA1 contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA (1), leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-shRNA1 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, thiselement creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (3). In addition to lentiviral elements, pLVX-shRNA1 contains a puromycin resistance gene (Puror)under the control of the murine phosphoglycerate kinase promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria. 应用pLVX-shRNA1 is available in the Lenti-X™ shRNA Expression System (Cat. No. 632177). Before it can be transduced into target cells, the vector must be cotransfected into 293T cells with our Lenti-X HTX Packaging System (Cat. Nos. 631247 and 631249) and packaged into viral particles. This packaging system allows the safe production of high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4). |
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武汉华尔纳生物科技有限公司,简称华尔纳生物(Warner Bio),坐 落于武汉生物产业基地·光谷生物城,是一家生物医学资源开发,科研生 产,产品销售集一体的科技型企业。
华尔纳生物(Warner Bio)始终专注于生命科学领域及细胞实验培养 方向,坚持为客户打造优质高效,安全可靠的生命科学产品,我们始终坚 持于以客户为中心、以品质求生存、以价值求发展、以服务求口碑的企业 理念,从而获得全国广大科研用户的一致认可。
团队以高效的服务品质,专业卓越的技术能力构建了华中地区最大的 生物细胞资源典藏中心,经过多年不懈的努力与发展,现与武汉大学、华 中科技大学、武汉协和医院、武汉同济医院、四川大学华西医院、首都医 科大学、中山大学附属第三医院等全国多所高校、医院、研究所单位达成 长期稳定的合作,同时为科研人员提供免费专业的售前售中售后服务,为 您的科研实验保驾护航。
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文献和实验产品技术背景 pRI系列载体是基于III类RNA聚合酶启动子:人类H1启动子的专用于哺乳动物细胞RNA干扰的载体。H1启动子在哺乳动物细胞内合成类似siRNA分子的小分子RNA。由于H1启动子有精确的转录起始位点和终止信号,H1启动子转录产物精确生成人工设计的shRNA,shRNA经过RISC剪切后形成有2个U突出末端的成熟siRNA。由于H1启动子对转录产物长度的严格限制,基本上杜绝了非特异性干扰片段的产生,将载体转染细胞后对其它基因的影响降到最低。 pRI系列载体已经成功用于多种
【1】 pSUPER vector http://www.oligoengine.com/pSUPER_New/pSUPER_Main2.html 【2】 pSilencer http://www.ambion.com/catalog/ProdGrps.html?fkApp=25&fkSubApp=145 【3】 psiRNA VECTOR http://www.invivogen.com/siRNA/psiRNA.htm 【4】 pGE-1 http://www
设计原则: 1.克隆到shRNA表达载体中的shRNA包括两个短反向重复序列,中间由一茎环(loop)序列分隔的,组成发夹结构,由polⅢ启动子控制。随后在连上5-6个T作为RNA聚合酶Ⅲ的转录终止子。 2.两个互补的寡核苷酸两端须带有限制性酶切位点。 3.Stratagene发现29个寡核苷酸较之原先推荐的23个寡核苷酸可以更有效的抑制目的基因。 4.在启动子下游的酶切位点下方紧连一个C,使插入片段和启动子有一定空间间隔以确保转录的发生。 5.ShRNA目的
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