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Sample-specific haplotype-resolved isoform characterization via long-read RNA-seq-based proteogenomics

Authors: Wissel, David; Sheynkman, Gloria; Robinson, Mark;

Sample-specific haplotype-resolved isoform characterization via long-read RNA-seq-based proteogenomics

Abstract

Bottom-up mass spectrometry (MS) supports disease research through the detection of protein isoforms. Recently, personalized proteomes have enabled more sensitive MS searches. Long-read RNA-seq (lrRNA-seq) data can be leveraged to create a sample-specific proteome in proteogenomics approaches to integrate genomic variation and alternative splicing events. We benchmark several algorithms for variant phasing on PacBio lrRNA-seq data and show that incorporating lrRNA-seq-based phased variants can increase peptide and protein isoform detection within MS-based searches. For this purpose, we develop a pipeline that constructs haplotype-resolved sample-specific proteomes, followed by MS search and annotation. Our workflow can be applied to samples containing matched MS and lrRNA-seq. We apply our workflow on a WTC11 sample and a ten-day osteoblast differentiation, highlighting the applicability of our work for both singular samples and more complicated experimental designs. We show that searching against sample-specific haplotype-resolved proteomes enables better detection and characterization of protein isoforms and supports the detection of linked variants. Consistent with previous work, genetic variation was consistently a much greater contributor to proteomic complexity than alternative splicing in our considered WTC11 sample. Our open-source Snakemake pipeline strives to support research and applications of haplotype-resolved MS searching based on lrRNA-seq data.

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    influence
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
1
Average
Average
Average
Green