Get precise genomic integration of your expression cassette
Use the PrecisionX™ Gene Knock-out HR Targeting Vector (MCS1-EF1α-RFP-T2A-Puro-pA-MCS2) to knock-out any gene or edit the genome. Clone your homology arms into MCS1 and MCS2, and use puromycin selection and RFP-positive imaging to find integrants. After you’ve identified clones with your gene-of-interest knocked-out or edited, you can remove the selection cassette using the Cre-LoxP system (learn more about Cre-LoxP excision here).
Why use an HR targeting vector?
Even though gene knock-outs can result from DSBs caused by Cas9 alone, SBI recommends the use of HR targeting vectors (also called HR donor vectors) for more efficient and precise mutation. HR donors can supply elements for positive or negative selection ensuring easier identification of successful mutation events. In addition, HR donors can include up to 6-8 kb of open reading frame for gene knock-ins or tagging, and, when small mutations are included in either 5’ or 3’ homology arms, can make specific, targeted gene edits.
Choose the right HR Targeting Vector for your project
Catalog # | HR Donor Vector | Features* | Application | |||
---|---|---|---|---|---|---|
Gene | Gene | Gene Edits | Gene Tagging | |||
HR100PA-1 | MCS1-LoxP-MCS2-MCS3-pA-LoxP-MCS4 | Basic HR Donor | ||||
HR110PA-1 | MCS1-EF1α-RFP-T2A-Puro-pA-MCS2 | Removable RFP marker and puromycin selection | ||||
HR120PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSPuro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP marker and puromycin selection | ||||
HR130PA-1 | T2A-GFP-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCSA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Co-express GFP with “tagged” gene via T2A Removable RFP marker and puromycin selection | ||||
HR150PA-1 | GFP-T2A-Luc-pA-loxP-EF1α-RFP-T2A-Puro-pA-LoxP-MCS | Tag with GFP fusion and co-express luciferase via T2A Removable RFP marker and puromycin selection | ||||
HR180PA-1 | IRES-GFP-pA-loxP-MCS1-EF1α-RFP-T2A-Puro-pA-LoxP-MCS2 | Co-express GFP with “tagged” gene via IRES Removable RFP marker and puromycin selection | ||||
HR210PA-1 | MCS1-LoxP-EF1α-GFP-T2A-Puro-P2A-hsvTK-pA-LoxP-MCS2 | Removable GFP marker, puromycin selection, and TK selection | ||||
HR220PA-1 | GFP-pA-LoxP-EF1α-RFP-T2A-Hygro-pA-LoxP-MCS | Tag with GFP fusion Removable RFP ,arker and hygromycin Selection | ||||
HR410PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2 | Removable GFP marker and puromycin selection | ||||
HR510PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2 | Removable RFP marker and hygromycin selection | ||||
HR700PA-1 | MCS1-EF1α-GFP-T2A-Puro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable GFP marker and puromycin selection | ||||
HR710PA-1 | MCS1-EF1α-RFP-T2A-Hygro-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable RFP marker and hygromycin selection | ||||
HR720PA-1 | MCS1-EF1α-Blasticidin-pA-MCS2-PGK-hsvTK | Enrich for on-target integration with negative TK selection** Removable blasticidin selection | ||||
GE602A-1 | pAAVS1D-PGK-MCS-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor | ||||
GE603A-1 | pAAVS1D-CMV-RFP-EF1α-copGFPpuro | First generation AAVS1-targeting HR Donor (positive control for GE602A-1) | ||||
GE620A-1 | AAVS1-SA-puro-MCS | Second generation AAVS1-targeting HR Donor Promoterless to knock-in any gene or promoter-gene combination | ||||
GE622A-1 | AAVS1-SA-puro-EF1α-MCS | Second generation AAVS1-targeting HR Donor Constitutive expression of your gene-of-interest | ||||
GE624A-1 | AAVS1-SA-puro-MCS-GFP | Second generation AAVS1-targeting HR Donor Create reporter cell lines | ||||
CAS620A-1 | AAVS1-SA-puro-EF1α-hspCas9 | Knock-in Cas9 to the AAVS1 site | ||||
MCS1-5"PB TR-EF1α-GFP-T2A-Puro-T2A-hsvTK-pA-3" PB TR-MCS2 | Use with the PiggyBac Transposon System Enables seamless gene editing with no residual footprint (i.e. completely remove vector sequences) | |||||
*All HR Target Vectors except PBHR100A-1 contain LoxP sites. Any sequences that are integrated between the two LoxP sites can be removed through transient expression of Cre Recombinase. **The clever design of these HR Donors enables enrichment for on-target integration events. A PGK-hsvTK cassette is included outside of the homology arms. Because of this configuration, on-target integration that results from homologous recombination will not include the PGK-hsvTK cassette—only randomly-integrated off-target events will lead to integration of PGK-hsvTK and resulting TK activity. Therefore, TK selection will negatively select against off-target integrants. Click on any one of these vectors to see a diagram of how the negative selection works. |
SystemBiosciences,简称SBI,美国加利福尼亚湾区新成立的生技公司,致力于独特,创新生物技术之开发,以研发利于基因及蛋白质功能鉴定,研究之崭新方法和工具为宗旨。
美国SBI代理SystemBiosciences,简称SBI,美国加州湾区新成立的生技公司,致力于独特、创新生物技术之开发,以研发利于基因及蛋白质功能鉴定、研究之崭新方法和工具为宗旨。现阶段研发重心为RNA干扰(RNAi)研究之相关工具。
现阶段研发重心为RNA干扰(RNAi)的研究之相关工具。系统Biosciences公司(SBI)致力于开发独特,革新的技术,为客户研究蛋白组学和基因组学功能提供研究工具.SBI是专业的慢病毒产品公司,提供基于慢病毒的所有相关产品,质粒,试剂盒及相关配套试剂和慢病毒扩展产品,如IPS细胞多功能性诱导试剂盒和RNAi筛选文库。System Biosciences继续创造新的独特产品以及改善我们完善的产品线。我们对提供领先技术的承诺与我们所有研究试剂和研究项目服务的高质量制造和质量控制相匹配。 2009年要寻找的东西:以下是即将推出的新产品。新型miRZips™:基于慢病毒载体的新型技术可永久敲低MicroRNA。双标记shRNA表达载体:SBI将发布新的功能强大的shRNA表达慢病毒载体pGreenPuro™,以便为稳定的RNAi实验选择稳定转导的细胞的GFP和Puro标记。甾醇反应pGreenFire™慢病毒报告子:基于慢病毒载体的转录报告子,用于监测与固醇感应转录因子相关的转录网络活性-心血管疾病途径的关键。诱导型表达慢载体:新的构建体将允许CDNA,shRNA和microRNA的“按需”表达。2007–2008年的新产品新的miR-SNaRE:MicroRNA小型非编码RNA富集系统,该系统使用带有表位标签的microRNA加工因子来提取蛋白质及其相关RNA。识别驱动RISC复合体的信使RNA。新的GeneNet™聚焦的shRNA库:这些聚焦的shRNA库编码一组针对所有与人类激酶,磷酸酶或细胞凋亡相关基因有关的特定功能或类别基因设计的shRNA集合。这些文库可进行有针对性的高通量RNAi筛选。新的miRNomeMicroRNA分析仪:qPCR阵列在预先格式化的板中包括microRNA分析,可用于人类的完全互补或小鼠单个microRNA的完全互补,每块板上带有三个内源参考RNA对照。所有基于SangermiRBasemicroRNA数据库的microRNA分析均已注册。新的Lenti-miR™MicroRNA前体克隆:在基于HIV的慢病毒载体中可获得更多的microRNA前体集合。超过580个单独的microRNA前体克隆可用阵列形式或合并的慢病毒形式用于HT筛选。新的基于慢病毒的干细胞报道者:SBI越来越多的慢病毒载体已被开发为将基因构建体在体内外几乎传递给任何细胞类型的最有效方法。SBI已将我们的慢病毒构建体系列扩展为干细胞报道分子。使用连接到GFP报告基因的细胞和途径特异性启动子,轻松创建转基因细胞系以监测细胞分化。QuantiMir™RT试剂盒:这项流行的新技术可通过一次cDNA合成同时进行实时qPCR定量分析数百种microRNA。设计您自己的microRNA测定法以进行创新性实验。癌症microRNAqPCR分析小组(OncoMir系列):预格式化的microRNA分析小组,用于评估95种已知与癌症有关的microRNA。干细胞microRNA分析小组:介绍了95种参与干细胞分化的microRNA,可同时监测干细胞的自我维持,造血途径和神经分化。SBI完善的产品线FullSpectrum™完整信使RNA扩增试剂盒:利用针对mRNA最常见基序的mRNA特异性引物提供完整的mRNA转录物(5"和3"末端)的无偏见,完整代表。GeneNet™siRNA库:全基因组,即用型,预包装的慢病毒库为筛选与生物学反应相关的基因功能提供了令人兴奋的机会。干扰素反应检测试剂盒:区分真正的RNA干扰和压力相关的细胞反应。PathNet™转录报告基因慢病毒载体:一种独特的方法,可创建各种稳定的报告基因细胞系,用于信号通路的研究。我们感谢您过去的支持,并期待为您提供最佳的新试剂,技术和服务,以加速您成功的研究目标。
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