Discipline of Intelligent Instrument and Equipment, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
Institute of Molecular Medicine, Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, China.
Lab Chip. 2024 May 14;24(10):2601-2621. doi: 10.1039/d4lc00105b.
Cells are the fundamental building blocks of living systems, exhibiting significant heterogeneity. The transcriptome connects the cellular genotype and phenotype, and profiling single-cell transcriptomes is critical for uncovering distinct cell types, states, and the interplay between cells in development, health, and disease. Nevertheless, single-cell transcriptome analysis faces daunting challenges due to the low abundance and diverse nature of RNAs in individual cells, as well as their heterogeneous expression. The advent and continuous advancements of next-generation sequencing (NGS) and third-generation sequencing (TGS) technologies have solved these problems and facilitated the high-throughput, sensitive, full-length, and rapid profiling of single-cell RNAs. In this review, we provide a broad introduction to current methodologies for single-cell transcriptome sequencing. First, state-of-the-art advancements in high-throughput and full-length single-cell RNA sequencing (scRNA-seq) platforms using NGS are reviewed. Next, TGS-based long-read scRNA-seq methods are summarized. Finally, a brief conclusion and perspectives for comprehensive single-cell transcriptome analysis are discussed.
细胞是生命系统的基本组成单位，表现出显著的异质性。转录组将细胞的基因型和表型联系起来，对单细胞转录组进行分析对于揭示不同的细胞类型、状态以及在发育、健康和疾病中细胞之间的相互作用至关重要。然而，由于单个细胞中 RNA 的丰度低且种类多样，以及其异质性表达，单细胞转录组分析面临着艰巨的挑战。新一代测序（NGS）和第三代测序（TGS）技术的出现和不断发展解决了这些问题，促进了高通量、敏感、全长和快速的单细胞 RNA 分析。在这篇综述中，我们对当前的单细胞转录组测序方法进行了广泛的介绍。首先，我们回顾了使用 NGS 的高通量和全长单细胞 RNA 测序（scRNA-seq）平台的最新进展。接下来，我们总结了基于 TGS 的长读 scRNA-seq 方法。最后，我们对全面的单细胞转录组分析进行了简要的总结和展望。
