Lindeman Ida, Stubbington Michael J T
Wellcome Sanger Institute, Hinxton, Cambridge, UK.
KG Jebsen Coeliac Disease Research Centre and Department of Immunology, University of Oslo, Oslo, Norway.
Methods Mol Biol. 2019;1935:223-249. doi: 10.1007/978-1-4939-9057-3_15.
In this chapter, we describe TraCeR and BraCeR, our computational tools for reconstruction of paired full-length antigen receptor sequences and clonality inference from single-cell RNA-seq (scRNA-seq) data. In brief, TraCeR reconstructs T-cell receptor (TCR) sequences from scRNA-seq data by extracting sequencing reads derived from TCRs by aligning the reads from each cell against synthetic TCR sequences. TCR-derived reads are then assembled into full-length recombined TCR sequences. BraCeR builds on the TraCeR pipeline and accounts for somatic hypermutations (SHM) and isotype switching. Here we discuss experimental design, use of the tools, and interpretation of the results.
在本章中,我们描述了TraCeR和BraCeR,这是我们用于从单细胞RNA测序(scRNA-seq)数据中重建配对全长抗原受体序列并进行克隆性推断的计算工具。简而言之,TraCeR通过将每个细胞的测序读数与合成T细胞受体(TCR)序列进行比对,从scRNA-seq数据中提取源自TCR的测序读数,从而重建TCR序列。然后将源自TCR的读数组装成全长重组TCR序列。BraCeR基于TraCeR流程构建,并考虑了体细胞超突变(SHM)和同种型转换。在这里,我们讨论实验设计、工具的使用以及结果的解释。