The SAM system involves three lentivirus plamids.Lenti sgRNA(MS2) purohave BsmBI restriction enzyme site, which is for insertion of the oligo sgRNA under the hU6 promoter. The result sgRNA(MS2) lentivirus express the fused MS2 and speci...
1.1.1主要材料 人胃癌细胞AGS由军事医学研究院辐射医学研究所保存;慢病毒表达载体Lenti-sgRNA(MS2)购自美国Addgene公司;血红蛋白(hemoglobin,HBG)sgRNA表达载体Lenti-sgHBG和肌联蛋白(titin,TTN)sgRNA表达载体Lenti-sgTTN由军事医学研究院辐射医学研究所构建并保存;SAM慢病毒激活载体dCas9-VP64和MS2-P65-HSF1购自上海...
To date, the most efficient dCas9-based activator is the Synergistic Activation Mediator (SAM) system whereby transcription activation domains are directly fused to dCas9 as well as tethered through MS2 loops engineered into the gRNA. Here, we show that dCas9 fused to the catalytic domain of ...
MS2 coat protein fused to the CBP HAT domain (MCP-CBP) where two MCP dimers recognize two MS2 loops in the gRNA and dCas9 thereby brings four CBP domains to the locus, dCas9-VPR where three
2. Insert the SgRNA oligo into lenti sgRNA(MS2)_puro plasmid, the map of lenti sgRNA(MS2)_puro is as follows: (1). Oligo annealing Mix the components above and anneal in a thermal cycler with the following conditions: On thermal cycler with heat lid. ...