High-throughput RNA isoform sequencing using programmed cDNA concatenation,Nature Biotechnology

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High-throughput RNA isoform sequencing using programmed cDNA concatenation
Nature Biotechnology ( IF 33.1 ) Pub Date : 2023-06-08 , DOI: 10.1038/s41587-023-01815-7
Aziz M Al'Khafaji ₁ , Jonathan T Smith ₁ , Kiran V Garimella ₁ , Mehrtash Babadi ₁ , Victoria Popic ₁ , Moshe Sade-Feldman _{1,

2} , Michael Gatzen ₁ , Siranush Sarkizova ₁ , Marc A Schwartz _{1,

3,

4,

5} , Emily M Blaum _{1,

2} , Allyson Day ₁ , Maura Costello ₁ , Tera Bowers ₁ , Stacey Gabriel ₁ , Eric Banks ₁ , Anthony A Philippakis ₁ , Genevieve M Boland ₆ , Paul C Blainey _{1,

7,

8} , Nir Hacohen _{1,

2,

9,

10}

Affiliation

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.

中文翻译：

使用程序化 cDNA 串联进行高通量 RNA 同工型测序

使用长读长技术的全长 RNA 测序方法可以捕获完整的转录本亚型，但其通量有限。我们引入了多重阵列同工型测序 (MAS-ISO-seq)，这是一种以可编程方式将互补 DNA (cDNA) 连接到最适合长读长测序的分子中的技术，可将通量提高 15 倍以上，每次运行可读取近 4000 万个 cDNA 读长。 Sequel IIe 测序仪。当应用于肿瘤浸润 T 细胞的单细胞 RNA 测序时，MAS-ISO-seq 发现差异剪接基因的发现增加了 12 至 32 倍。

更新日期：2023-06-08

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