| 项目编号 |
PRJCA003458 |
| 项目标题 |
HIT-scISOseq: High-throughput and High-accuracy Single-cell Full-length Isoform Sequencing for Corneal Epithelium |
| 涉及领域 |
Model organism |
| 数据类型 |
Transcriptome or Gene expression
|
| 物种名称 |
Mus musculus
Macaca fascicularis
Homo sapiens
|
| 描述信息 |
Single-cell isoform sequencing can reveal transcriptomic dynamics in individual cells invisible to bulk- and single-cell RNA analysis based on short-read sequencing. However, current long-read single-cell sequencing technologies have been limited by low throughput and high error rate. Here we introduce HIT-scISOseq for high-throughput single-cell isoform sequencing. This method was made possible by full-length cDNA capture using biotinylated PCR primers, and by our novel library preparation procedure that combines head-to-tail concatemeric full-length cDNAs into a long SMRTbell insert for high-accuracy PacBio sequencing. HIT-scISOseq yields > 10 million high-accuracy full-length isoforms in a single PacBio Sequel II 8M SMRT Cell, providing > 8 times more data output than the standard single-cell isoform PacBio sequencing protocol. We exemplified HIT-scISOseq by first studying transcriptome profiles of 4,000 normal and 8,000 injured corneal epitheliums from cynomolgus monkeys. We constructed dynamic transcriptome landscapes of known and rare cell types, revealed novel isoforms, and identified injury-related splicing and switching events that are previously not accessible with low throughput isoform sequencing. HIT-scISOseq represents a high-throughput, cost-effective, and technically simple method to accelerate the burgeoning field of long-read single-cell transcriptomics. |
| 样品范围 |
Single cell |
| 发布日期 |
2023-03-28 |
| 出版信息 |
| PubMed ID |
文章标题 |
杂志名称 |
Doi |
发表年份 |
|
|
HIT-scISOseq: High-throughput and High-accuracy Single-cell Full-length Isoform Sequencing
|
Research Square
|
10.21203/rs.3.rs-114035/v1
|
2022
|
| 37149708
|
High-throughput and high-accuracy single-cell RNA isoform analysis using PacBio circular consensus sequencing
|
Nature Communications
|
10.1038/s41467-023-38324-9
|
2023
|
|
|
| 项目资金来源 |
| 机构 |
项目类型 |
授权项目ID |
授权项目名称 |
| National Natural Science Foundation of China (NSFC)
|
|
81722034
|
|
|
|
| 提交者 |
Chuanle
Xiao (xiaochuanle@126.com)
|
| 提交单位 |
Zhongshan Ophthalmic Center,Sun Yat-sen University |
| 提交日期 |
2020-05-31 |
