High-throughput RNA isoform sequencing using programmed cDNA concatenation

被引：76

作者：

Al'Khafaji, Aziz M. ^{[1
]}

Smith, Jonathan T. ^{[1
]}

Garimella, Kiran V. ^{[1
]}

Babadi, Mehrtash ^{[1
]}

Popic, Victoria ^{[1
]}

Sade-Feldman, Moshe ^{[1
,2
]}

Gatzen, Michael ^{[1
]}

Sarkizova, Siranush ^{[1
]}

Schwartz, Marc A. ^{[1
,3
,4
,5
]}

Blaum, Emily M. ^{[1
,2
]}

Day, Allyson ^{[1
]}

Costello, Maura ^{[1
]}

Bowers, Tera ^{[1
]}

Gabriel, Stacey ^{[1
]}

Banks, Eric ^{[1
]}

Philippakis, Anthony A. ^{[1
]}

Boland, Genevieve M. ^{[6
]}

Blainey, Paul C. ^{[1
,7
,8
]}

Hacohen, Nir ^{[1
,2
,9
,10
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge 02142, MA USA

[2] Massachusetts Gen Hosp, Ctr Canc Res, Dept Med, Boston, MA 02114 USA

[3] Harvard Med Sch, Dept Pediat, Boston, MA USA

[4] Boston Childrens Hosp, Div Hematol Oncol, Boston, MA USA

[5] Dana Farber Canc Inst, Dept Pediat Oncol, Boston, MA USA

[6] Harvard Med Sch, Massachusetts Gen Hosp, Div Surg Oncol, Boston, MA USA

[7] MIT, Dept Biol Engn, Cambridge, MA 02139 USA

[8] MIT, Koch Inst Integrat Canc Res, Cambridge, MA 02139 USA

[9] Harvard Med Sch, Boston, MA 02115 USA

[10] Massachusetts Gen Hosp, Ctr Immunol & Inflammatory Dis, Charlestown, MA 02129 USA

来源：

NATURE BIOTECHNOLOGY | 2024年 / 42卷 / 04期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1038/s41587-023-01815-7

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Full-length RNA-sequencing methods using long-read technologies can capture complete transcript isoforms, but their throughput is limited. We introduce multiplexed arrays isoform sequencing (MAS-ISO-seq), a technique for programmably concatenating complementary DNAs (cDNAs) into molecules optimal for long-read sequencing, increasing the throughput >15-fold to nearly 40 million cDNA reads per run on the Sequel IIe sequencer. When applied to single-cell RNA sequencing of tumor-infiltrating T cells, MAS-ISO-seq demonstrated a 12- to 32-fold increase in the discovery of differentially spliced genes. Programmable concatenation of cDNA molecules increases the throughput of PacBio sequencing about 15-fold.

引用

页码：582 / 586

页数：16

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