Adaptive nanopore sequencing for single cell characterization of cancer mutations and gene rearrangements

Autor: Susan M. Grimes, Heon Seok Kim, Sharmili Roy, Anuja Sathe, Carlos I. Ayala, Alison F. Almeda-Notestine, Sarah Haebe, Tanaya Shree, Ronald Levy, Billy T. Lau, Hanlee P. Ji
Rok vydání: 2022
Popis: In this proof-of-concept study, we developed a single cell method that identifies somatic alterations found in coding regions of mRNAs and integrates these mutation genotypes with their matching cell transcriptomes. We used nanopore adaptive sampling on single cell cDNA libraries, generated long read sequences from target gene transcripts and identified coding variants among individual cells. Short-read single cell transcriptomes characterized the cell types with mutations. We delineated CRISPR edits from a cancer cell line. From primary cancer samples, we targeted hundreds of cancer genes, identified somatic coding mutations and a gene rearrangement among individual tumor cells.
Databáze: OpenAIRE