Single-cell long-read sequencing of the experience-induced transcriptome
Cavalier, S.; Hook, P. W.; Huganir, R. L.; Timp, W.
Show abstract
Neural activity drives transcriptional events that are critical for learning. Activity-induced transcript isoform expression and alternative splicing are cell-type specific events typically obscured by sequencing approaches that restrict read length. We combined single-cell transcriptomics with Nanopore long-read sequencing to resolve these phenomena, generating the first dataset profiling learning-induced gene and isoform expression in individual mouse hippocampal cells. Due to their length, the majority of reads we generated could be uniquely mapped to their respective gene and isoform features in the mouse reference. [~]20k cells from 16 mouse samples were sequenced and clustered on the basis of gene expression, yielding 21 hippocampal cell types. Differential expression analysis revealed 1,266 significantly experience-variable isoforms, revealing novel splicing behavior in many synaptic genes. This work is the first to comprehensively profile activity-induced isoform expression, demonstrating that single cell long-read sequencing can reveal new layers of transcriptional complexity.
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