A linear-amplification VDJ-seq technique for quantification of immunoglobulin and T cell receptor diversity
A linear-amplification VDJ-seq technique for quantification of immunoglobulin and T cell receptor diversity
复制标题
用于定量免疫球蛋白和 T 细胞受体多样性的线性扩增 VDJ-seq 技术
DOI：
10.1139/gen-2019-0096
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发表时间：
2020
期刊：
影响因子：
3.1
通讯作者：
Hao Bingtao
中科院分区：
文献类型：
--
作者：
Zhao Hao;Li Zhaoqiang;Zhu Yongchang;Hao Bingtao
The V(D)J recombination is essential for generating a highly diverse repertoire of antigen receptors expressed on T and B lymphocytes. Here, we developed a linear-amplification VDJ-seq technique for quantifying V(D)J recombination of antigen receptor genes. This technique takes advantage of linear amplification using in vitro transcription and reverse transcription to avoid bias generated by the PCR amplification of low copy number of target DNA. The unrearranged alleles are removed by in vitro cleavage with the CRISPR-Cas9 system. The linear-amplification VDJ-seq assay was applied in quantification of the Vκ-Jκ recombination of the mouseIgκgene with Jκ capture primers. The Jκ genes were detected in 95.86% of clean reads with more than half containing the Vκ gene, indicating high specificity of capturing and amplification. We also applied this approach to quantify the usage of Jα within the Trav12 gene family of theTcragene.
