Second-Strand Synthesis-Based Massively Parallel scRNA-Seq Reveals Cellular States and Molecular Features of Human Inflammatory Skin Pathologies.
Second-Strand Synthesis-Based Massively Parallel scRNA-Seq Reveals Cellular States and Molecular Features of Human Inflammatory Skin Pathologies.
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基于第二链合成的大规模平行单细胞RNA测序揭示人类炎症性皮肤疾病的细胞状态和分子特征
DOI：
10.1016/j.immuni.2020.09.015
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发表时间：
2020-10-13
期刊：
影响因子：
32.4
通讯作者：
Shalek AK
中科院分区：
文献类型：
--
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
高通量单细胞RNA测序（scRNA - seq）方法通过增加可同时分析的细胞数量，能够对复杂生物样本进行表征。然而，与低通量策略相比，这些方法从每个细胞中获取的信息较少。为了准确报告细胞关键表型特征的表达，需要既高保真又高通量的scRNA - seq平台。为了满足这一需求，我们创建了Seq - Well S3（“第二链合成”），这是一种大规模并行的scRNA - seq方案，它使用随机引物的第二链合成来恢复已成功逆转录但由于模板转换效率低下而未添加后续全转录组扩增所需的第二个寡核苷酸接头的互补DNA（cDNA）分子。与之前的版本相比，Seq - Well S3将转录本捕获效率和基因检测效率分别提高了多达10倍和5倍。我们使用Seq - Well S3绘制了五种人类炎症性皮肤病的转录图谱，从而为进一步研究人类皮肤炎症提供了资源。
Seq - Well S3利用第二链合成来提高转录本捕获效率
Seq - Well S3与一流的商业平台进行了基准比较
Seq - Well S3被应用于分析皮肤病中的炎症细胞状态
对皮肤炎症的分析揭示了独特且保守的细胞表型
休斯等人报道了一种高通量单细胞RNA测序技术“Seq - Well S3”的开发，该技术能够提高灵敏度并改进对包括转录因子、细胞因子和细胞因子受体在内的基因的检测。作者使用Seq - Well S3定义了多种皮肤病中的炎症细胞状态。
High-throughput single-cell RNA-sequencing (scRNA-seq) methodologies enable characterization of complex biological samples by increasing the number of cells that can be profiled contemporaneously. Nevertheless, these approaches recover less information per cell than low-throughput strategies. To accurately report the expression of key phenotypic features of cells, scRNA-seq platforms are needed that are both high fidelity and high throughput. To address this need, we created Seq-Well S3 (“Second-Strand Synthesis”), a massively parallel scRNA-seq protocol that uses a randomly primed second-strand synthesis to recover complementary DNA (cDNA) molecules that were successfully reverse transcribed but to which a second oligonucleotide handle, necessary for subsequent whole transcriptome amplification, was not appended due to inefficient template switching. Seq-Well S3 increased the efficiency of transcript capture and gene detection compared with that of previous iterations by up to 10- and 5-fold, respectively. We used Seq-Well S3 to chart the transcriptional landscape of five human inflammatory skin diseases, thus providing a resource for the further study of human skin inflammation. Seq-Well S3 uses second-strand synthesis to improve transcript capture Seq-Well S3 was benchmarked against a best-in-class commercial platform Seq-Well S3 was applied to profile inflammatory cell states in skin diseases Analysis of skin inflammation uncovered unique and conserved cellular phenotypes Hughes et al. report the development of a technique for high-throughput single-cell RNA-sequencing, “Seq-Well S3,” that enables increased sensitivity and improved detection of genes including transcription factors, cytokines, and cytokine receptors. Using Seq-Well S3, the authors define inflammatory cell states across multiple skin diseases.
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影响因子：
30.5
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影响因子：
32.4
作者：
Guilliams M;Dutertre CA;Scott CL;McGovern N;Sichien D;Chakarov S;Van Gassen S;Chen J;Poidinger M;De Prijck S;Tavernier SJ;Low I;Irac SE;Mattar CN;Sumatoh HR;Low GHL;Chung TJK;Chan DKH;Tan KK;Hon TLK;Fossum E;Bogen B;Choolani M;Chan JKY;Larbi A;Luche H;Henri S;Saeys Y;Newell EW;Lambrecht BN;Malissen B;Ginhoux F
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Ginhoux F
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82.9
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影响因子：
64.8
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影响因子：
32.4
作者：
Dutertre, Charles-Antoine;Becht, Etienne;Ginhoux, Florent
通讯作者：
Ginhoux, Florent