将这些TCR转导到从外周血单个核细胞(PBMCs)中收集的同种异体T细胞中,为每个患者生成20个TCR-T细胞系。从每个患者的肿瘤中选择高到中等mRNA水平的100个突变位点作为潜在的肿瘤抗原,并通过WES和bulk RNA-seq检测这些位点。构建5个串联基因并将其转导到同种异体淋巴母细胞瘤细胞系(LCLs)中。 实验结果: 将每个TCR-T细...
5’RACE利用RNA,只使用一对引物对靶向TCR链的恒定区域和mRNA5’端,这消除了多重PCR所带来的偏差,并允许捕获给定样品中存在的所有TCR。 2 bulk测序和单细胞测序 关于测序方法,bulk测序或单细胞测序可用于分析T细胞库。 bulk测序: 它对样本中未配对的TCR链的聚集进行测序,通常用于分析大规模TCR多样性并比较患者队列...
第二种TCR测序方法是基于5’ RACE,这种方法需要使用具有末端转移酶活性的逆转录酶反转录RNA,从而在cDNA的3’ 端加入非模板C寡核苷酸,用于下游的cDNA扩增,非模板C寡核苷酸叫做模板转换寡核苷酸(Template switch oligo,TSO)。在第一链合成过程中,当到达 RNA 模板的 5' 末端时,逆转录酶的末端转移酶活性会在新合成的...
detailed assessment of neoTCR-characteristics affecting therapeutic efficacy is mostly missing. Previously, we identified diverse neoTCRs restricted to different neoantigens in a melanoma patient. In this work, we now combine single-cell TCR-sequencing and RNA-sequencing after neoantigen...
Methods We performed single-cell RNA sequencing (scRNA-Seq) and T cell receptor sequencing (scTCR-Seq) on CD8+ T cells infiltrating YTN2 and YTN16 tumors. Subsequently, we cloned five and four TCRs from YTN2 and YTN16, respectively, based on scTCR-Seq data. The cloned TCRs were ...
5’RACE利用RNA,只使用一对引物对靶向TCR链的恒定区域和mRNA5’端,这消除了多重PCR所带来的偏差,并允许捕获给定样品中存在的所有TCR。 2 bulk测序和单细胞测序 关于测序方法,bulk测序或单细胞测序可用于分析T细胞库。 bulk测序: 它对样本中未配对的TCR链的聚集进行测序,通常用于分析大规模TCR多样性并比较患者队...
MTGDC: A multi-scale tensor graph diffusion clustering for single-cell RNA sequencing data IEEE/ACM Transactions on Computational Biology and Bioinformatics; IF: 4.500; DOI: 10.1109/TCBB.2023.3293112 内容概要:提出了一种名为MTGDC(https://github.com/lqmmring/MTGDC)的多尺度张量图扩散聚类方法,用于sc...
Single-cell RNA sequencing coupled to TCR profiling of large granular lymphocyte leukemia T cells 期刊:Nature communicationsCancer cell 影响因子:14.919 发表单位:Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA ...
Learn about our SMARTer approach to single T-cell sequencing that obtains alpha-beta pairing information.
Figure 3. TCR sequencing libraries with similar size distributions are produced from human and rhesus macaque RNA inputs.TRA/TRBmixed libraries were prepared using 100 ng total RNA isolated from PBMC of human (Panel A) or a rhesus macaque donor (Panel B). Complet...