Depletion of abundant transcripts using Cas-9 and long-read sequencing improves transcriptome annotation of the polar bear
Who's speaking at this Webinar?
Ashley L. Byrne
Graduate Student Researcher, University of California, Santa Cruz
Transcriptome studies evaluating whole blood and tissues are often confounded by overrepresentation of highly abundant transcripts, which compete with and prevent detection of rare RNA transcripts, obscuring their biological importance.
In this webinar, we will discuss Long-DASH, an adapted CRISPR/Cas9-based method, to deplete highly-abundant transcripts for isoform-level transcriptome analysis using long-read sequencing in a non-model organism.
We will demonstrate how we used Long-DASH to sequence full-length cDNA using Oxford Nanopore R2C2 long-read and Illumina Smart-seq2 approaches and applied the method to create an isoform-level transcriptome from whole blood samples derived from polar bears (Ursus maritimus).
Here viewers should expect to learn:
- How to deplete unwanted transcripts using Long-DASH
- About the R2C2 method and C3POa pipeline to improve accuracy
- How to identify isoforms using Mandalorion
- How to identify novel un-annotated isoforms using SQA