EMBL Australia Partner Laboratory Network at the Australian National University, Acton, Canberra, ACT, 2601, Australia.
Pompeu Fabra University (UPF), E08003, Barcelona, Spain.
Genome Biol. 2022 Jul 8;23(1):153. doi: 10.1186/s13059-022-02715-w.
Nanopore sequencing enables the efficient and unbiased measurement of transcriptomes. Current methods for transcript identification and quantification rely on mapping reads to a reference genome, which precludes the study of species with a partial or missing reference or the identification of disease-specific transcripts not readily identifiable from a reference. We present RATTLE, a tool to perform reference-free reconstruction and quantification of transcripts using only Nanopore reads. Using simulated data and experimental data from isoform spike-ins, human tissues, and cell lines, we show that RATTLE accurately determines transcript sequences and their abundances, and shows good scalability with the number of transcripts.
纳米孔测序能够高效且无偏地测量转录组。目前用于转录本鉴定和定量的方法依赖于将reads 映射到参考基因组,这排除了对部分或缺少参考基因组的物种的研究,或对难以从参考基因组鉴定的疾病特异性转录本的研究。我们提出了 RATTLE,这是一种仅使用纳米孔读取来执行无参考重建和转录本定量的工具。使用模拟数据和来自异构体插入物、人体组织和细胞系的实验数据,我们表明 RATTLE 可以准确地确定转录本序列及其丰度,并且随着转录本数量的增加具有良好的可扩展性。