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.

中文翻译：

---

基于先进测序的高通量和长读长单细胞转录组分析

细胞是生命系统的基本组成部分，表现出显着的异质性。转录组将细胞基因型和表型联系起来，单细胞转录组分析对于揭示不同的细胞类型、状态以及发育、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、性质多样以及它们的异质表达，单细胞转录组分析面临着严峻的挑战。新一代测序（NGS）和第三代测序（TGS）技术的出现和不断进步解决了这些问题，并促进了单细胞RNA的高通量、灵敏、全长和快速分析。在这篇综述中，我们广泛介绍了当前的单细胞转录组测序方法。首先，回顾了使用 NGS 的高通量全长单细胞 RNA 测序 (scRNA-seq) 平台的最新进展。接下来，总结了基于 TGS 的长读长 scRNA-seq 方法。最后，讨论了综合单细胞转录组分析的简要结论和展望。