Single-cell Iso-Sequencing enables rapid genome annotation for scRNAseq analysis. Issue 3 (10th February 2022)

• Record Type: Journal Article
• Authors: Healey, Hope M; Bassham, Susan; Cresko, William A
• Editors: Sanchez Alvarado, A
• Is Part Of: Genetics. Volume 220:Issue 3(2022)
• Publication Date: 2022-02-10
• Subjects: PacBio -- emerging model organisms -- 10X Genomics; Genetics -- Periodicals; 576.5
• DOI: 10.1093/genetics/iyac017
• Languages: English
• ISSNs: 0016-6731

Abstract

Abstract: Single-cell RNA sequencing is a powerful technique that continues to expand across various biological applications. However, incomplete 3′-UTR annotations can impede single-cell analysis resulting in genes that are partially or completely uncounted. Performing single-cell RNA sequencing with incomplete 3′-UTR annotations can hinder the identification of cell identities and gene expression patterns and lead to erroneous biological inferences. We demonstrate that performing single-cell isoform sequencing in tandem with single-cell RNA sequencing can rapidly improve 3′-UTR annotations. Using threespine stickleback fish ( Gasterosteus aculeatus ), we show that gene models resulting from a minimal embryonic single-cell isoform sequencing dataset retained 26.1% greater single-cell RNA sequencing reads than gene models from Ensembl alone. Furthermore, pooling our single-cell sequencing isoforms with a previously published adult bulk Iso-Seq dataset from stickleback, and merging the annotation with the Ensembl gene models, resulted in a marginal improvement (+0.8%) over the single-cell isoform sequencing only dataset. In addition, isoforms identified by single-cell isoform sequencing included thousands of new splicing variants. The improved gene models obtained using single-cell isoform sequencing led to successful identification of cell types and increased the reads identified of many genes in our single-cell RNA sequencing stickleback dataset. Our work illuminates single-cell isoform sequencing as a cost-effective and efficient mechanism to rapidly annotate genomes for single-cell RNA sequencing.