Oh, S. A. et al. (2022). High-efficiency nonviral CRISPR/Cas9-mediated gene editing of human T cells using plasmid donor DNA. J. Exp. Med. 219, e20211530. 在抗体圈微信公众号回复“JPM23”可下载60家药企PPT合集
plasmid/bacterial cells retroviral vector/mammalian cells In vivo display methods include cell-surface display methods in which a plasmid is introduced into the host cell encoding a fusion protein consisting of the protein or polypeptide of interest fused to a cell surface protein or polypeptide. The...
For recovery of TCR sequences, genomic DNA (obtained using the DNeasy Blood & Tissue Kit according to manufacturer’s protocol) or TCR plasmid library was used as template for PCR. The PCR was performed using a forward primer annealing to a 5’ upstream plasmid sequence and a reverse primer...
The invention discloses a recombinant plasmid relevant to the EB virus characteristic TCR gene and a method for reverting normal T cell to obtain the anti-EBV characteristic CTL. The recombinant plasmid consists of TCRV Alpha 15 gene, pIRES vector and TCR V Beta 1gene; the TCRV Alpha 15-p...
HEK-293T cells were seeded in a 10 cm tissue culture dish at a density of 5 × 106cells per dish. The next day, HEK-293T cells were transfected with the TCR expressing plasmid and packaging plasmids (pMDLg/pRRE, pRSV-Rev, pMD2.G) using polyethylenimine (Polyplus, cat. PT-115...
and then attached to a human antibody Fc region. The single-chain TCRα-linker-TCRβ-Fc insert was cloned into a pcDNA vector (pcDNA/scTCR-Fc) (Supplementary Table7). The scTCR-Fc plasmid was transfected into the HEK293F cells (Thermo Fisher, Cat: R79007), and the soluble single-chai...
(Addgene plasmid # 52114;http://n2t.net/addgene:52114; RRID:Addgene_52114)), followed by a three-way ligation using Gibson Assembly Cloning (NEB, E2611S) at 50 °C for 15 min and stored on ice. The resulting pTCR-MIR expression vectors encoded TCRα and TCRβ fragments ...
然后将产物转化入e.colidh5αcompetentcells(购自takara),通过氨苄青霉素(ampicillin,amp)筛选出阳性菌落,挑取阳性菌落送武汉天一辉远生物科技有限公司测质粒全序列,将序列正确的菌落接种入250ml含amp的lb液体培养基中,于37℃和220rpm摇菌16至24h,使用endofreeplasmidgigakit(购自qiagen)从菌液中提取质粒,将质粒命名...
TCRplasmidlibraries.Usingsingle-celltranscriptomesequencingdata,informationonthe tumor-reactivitymarkersPDCD1,ENTPD1,andCXCL13wasobtained;highclonalTCR sequenceinformationwasobtainedusingsingle-cellTCRsequencingdatatoscreen potentialtumor-reactiveTCRsequencesforeachpatient.Targetplasmidswereselected fromthepre-preparedTCR...
质粒提取试剂盒QIAGEN Plasmid Midi Kit和胶回收试剂盒QIAEX II Gel Extraction Kit购自QIAGEN公司,Lipofection 3000购自ThermoFisher Scientific公司,BsmBⅠ-v2内切酶和T4 Ligase购自New England BioLabs公司,TCRP1抗体购自Novus Biologicals公司,...