High-throughput and accuracy of the next-generation sequencing (NGS) helped single-cell RNA sequencing (scRNA-seq) become one of the most widely used technologies for identification of cell types, cellular states and rare phenotypes. Despite the latest improvements of scRNA-seq, most current methods rely on poly(A) tail capture and thus are mainly restricted to protein-coding RNAs, while missing a substantial fraction of the non-coding transcriptome. Moreover, most of these methods are developed to be compatible with NGS and rely on 3’ end sequencing of RNA, which is limited to gene, but not its isoform, identification and in turn complicates further functional analyses of single cells. Therefore, third generation sequencing (long-read) based scRNA-seq technique, which enables to profile the whole transcriptome in full-length, could broaden the understanding about regulatory cellular processes and functions.
Within the PhD project’s scope, the candidate will develop full-length whole RNA-sequencing employing advanced, multi-step reaction compatible microfluidics systems and long-read sequencing technologies. The applicant will apply this method to investigate dynamics of RNA maturation and regulatory roles of non-coding RNAs during cell differentiation. Along the project development and implementation, the candidate will acquire multidisciplinary skills in microfluidics, biochemistry and bioinformatics.
Supervisor:
Dr. Simonas Juzėnas
Contacts: