Targeting individual cells by barcode in pooled sequence libraries.
Targeting individual cells by barcode in pooled sequence libraries.
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DOI：
10.1093/nar/gky856
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发表时间：
2019-01-10
影响因子：
14.9
通讯作者：
Blainey PC
中科院分区：
文献类型：
--
作者：
Ranu N;Villani AC;Hacohen N;Blainey PC
现在，RNA-seq并行的数千个单元的转录分析是常规的。但是，由于依赖汇总图书馆的准备，将分析靶向特定感兴趣的细胞是困难的。在这里，我们提出了一种多重PCR方法，用于从合并的单细胞序列文库中靶向测序。我们证明了这种分子富集方法对由原代人血细胞产生的汇总单细胞RNA-seq库中的多种细胞类型。我们展示了如何将分子富集与FACS结合起来，以有效地靶向超稀有细胞类型，例如最近确定的AXL+ SIGLEC6+树突状细胞（AS DC）子集，以减少所需的测序努力，以将所需的测序介绍为100--折叠。我们的结果表明，可以将合并测序文库中识别细胞的DNA条形码用作富集特定感兴趣分子的目标，例如从一组目标细胞中读取。
Transcriptional profiling of thousands of single cells in parallel by RNA-seq is now routine. However, due to reliance on pooled library preparation, targeting analysis to particular cells of interest is difficult. Here, we present a multiplexed PCR method for targeted sequencing of select cells from pooled single-cell sequence libraries. We demonstrated this molecular enrichment method on multiple cell types within pooled single-cell RNA-seq libraries produced from primary human blood cells. We show how molecular enrichment can be combined with FACS to efficiently target ultra-rare cell types, such as the recently identified AXL+SIGLEC6+ dendritic cell (AS DC) subset, in order to reduce the required sequencing effort to profile single cells by 100-fold. Our results demonstrate that DNA barcodes identifying cells within pooled sequencing libraries can be used as targets to enrich for specific molecules of interest, for example reads from a set of target cells.
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影响因子：
64.8
作者：
Shalek, Alex K.;Satija, Rahul;Shuga, Joe;Trombetta, John J.;Gennert, Dave;Lu, Diana;Chen, Peilin;Gertner, Rona S.;Gaublomme, Jellert T.;Yosef, Nir;Schwartz, Schraga;Fowler, Brian;Weaver, Suzanne;Wang, Jing;Wang, Xiaohui;Ding, Ruihua;Raychowdhury, Raktima;Friedman, Nir;Hacohen, Nir;Park, Hongkun;May, Andrew P.;Regev, Aviv
通讯作者：
Regev, Aviv
影响因子：
64.5
作者：
Enge M;Arda HE;Mignardi M;Beausang J;Bottino R;Kim SK;Quake SR
通讯作者：
Quake SR
影响因子：
48
作者：
Vitak SA;Torkenczy KA;Rosenkrantz JL;Fields AJ;Christiansen L;Wong MH;Carbone L;Steemers FJ;Adey A
通讯作者：
Adey A
DOI：
10.1126/science.1247651
发表时间：
2014-02-14
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Jaitin DA;Kenigsberg E;Keren-Shaul H;Elefant N;Paul F;Zaretsky I;Mildner A;Cohen N;Jung S;Tanay A;Amit I
通讯作者：
Amit I
单细胞RNA测序揭示了T辅助细胞合成从头开始的类固醇，从而有助于免疫稳态。
影响因子：
8.8
作者：
Mahata B;Zhang X;Kolodziejczyk AA;Proserpio V;Haim-Vilmovsky L;Taylor AE;Hebenstreit D;Dingler FA;Moignard V;Göttgens B;Arlt W;McKenzie AN;Teichmann SA
通讯作者：
Teichmann SA