TAS-Seq is a robust and sensitive amplification method for bead-based scRNA-seq.
TAS-Seq is a robust and sensitive amplification method for bead-based scRNA-seq.
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DOI：
10.1038/s42003-022-03536-0
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发表时间：
2022-06-27
影响因子：
5.9
通讯作者：
中科院分区：
文献类型：
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单细胞RNA测序（scRNA-seq）对于分析细胞异质性很有价值。细胞组成的准确性对于从scRNA-seq数据分析细胞-细胞相互作用网络至关重要。然而，基于液滴和平板的scRNA-seq技术具有细胞采样偏差，这可能会影响scRNA-seq数据集的细胞组成。在这里，我们开发了基于终止子、末端转移酶和基于微球/珠的scRNA-seq平台的scRNA-seq的终止子辅助固相cDNA扩增和测序（TAS-Seq）。TAS-Seq显示出对末端转移酶反应中的变化的高耐受性，这使得现有的基于末端转移酶的scRNA-seq方法的处理复杂化。在小鼠和人肺样本中，TAS-Seq产生的scRNA-seq数据与流式细胞术数据高度相关，显示出比10 X Chromium v2和Smart-seq 2更高的基因检测灵敏度和更稳健的重要细胞间相互作用检测以及细胞亚群中生长因子/白细胞介素的表达。扩大TAS-Seq的应用将提高对单细胞水平肺生物学的理解和图谱构建。终止子辅助的固相cDNA扩增和测序（TASseq）使用终止子、末端转移酶和基于珠的平台来改进单细胞RNA-seq文库的生成。
Single-cell RNA-sequencing (scRNA-seq) is valuable for analyzing cellular heterogeneity. Cell composition accuracy is critical for analyzing cell–cell interaction networks from scRNA-seq data. However, droplet- and plate-based scRNA-seq techniques have cell sampling bias that could affect the cell composition of scRNA-seq datasets. Here we developed terminator-assisted solid-phase cDNA amplification and sequencing (TAS-Seq) for scRNA-seq based on a terminator, terminal transferase, and nanowell/bead-based scRNA-seq platform. TAS-Seq showed high tolerance to variations in the terminal transferase reaction, which complicate the handling of existing terminal transferase-based scRNA-seq methods. In murine and human lung samples, TAS-Seq yielded scRNA-seq data that were highly correlated with flow-cytometric data, showing higher gene-detection sensitivity and more robust detection of important cell–cell interactions and expression of growth factors/interleukins in cell subsets than 10X Chromium v2 and Smart-seq2. Expanding TAS-Seq application will improve understanding and atlas construction of lung biology at the single-cell level. Terminator-assisted solid-phase cDNA amplification and sequencing (TASseq) uses a terminator, terminal transferase and bead-based platform to improve generation of single-cell RNA-seq libraries.
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DOI：
10.1146/annurev-pathol-020712-163930
发表时间：
2013-01-24
期刊：
Annual review of pathology
影响因子：
--
作者：
Duffield JS;Lupher M;Thannickal VJ;Wynn TA
通讯作者：
Wynn TA
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
14.9
作者：
Huang H;Goto M;Tsunoda H;Sun L;Taniguchi K;Matsunaga H;Kambara H
通讯作者：
Kambara H
影响因子：
32.4
作者：
Hughes TK;Wadsworth MH 2nd;Gierahn TM;Do T;Weiss D;Andrade PR;Ma F;de Andrade Silva BJ;Shao S;Tsoi LC;Ordovas-Montanes J;Gudjonsson JE;Modlin RL;Love JC;Shalek AK
通讯作者：
Shalek AK
影响因子：
64.5
作者：
Cho CS;Xi J;Si Y;Park SR;Hsu JE;Kim M;Jun G;Kang HM;Lee JH
通讯作者：
Lee JH