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Accession IconSRP099636

Analysis of C. elegans muscle transcriptome using trans-splicing-based RNA tagging (SRT)

Organism Icon Caenorhabditis elegans
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Technology Badge IconIllumina HiSeq 2500

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Description
Current approaches to profiling tissue-specific gene expression in C. elegans require delicate manipulation and are difficult under certain conditions, e.g. from dauer or aging worms. We have developed an easy and robust method for tissue-specific RNA-seq by taking advantage of the endogenous trans-splicing process. In this method, transgenic worms are generated in which a spliced leader (SL) RNA gene is fused with a sequence tag and driven by a tissue-specific promoter. Only in the tissue of interest, the tagged SL RNA gene is transcribed and then trans-spliced onto mRNAs. The tag allows enrichment and sequencing of mRNAs from that tissue only. As a proof of principle, we profiled the muscle transcriptome, which showed high coverage and efficient enrichment of muscle specific genes, with low background noise. To demonstrate the robustness of our method, we profiled muscle gene expression in dauer larvae and aging worms, revealing gene expression changes consistent with the physiology of these stages. The resulting muscle transcriptome also revealed 461 novel RNA transcripts, likely muscle-expressed long non-coding RNAs. In summary, the splicing-based RNA tagging (SRT) method provides a convenient and robust tool to profile trans-spliced genes and identify novel transcripts in a tissue-specific manner, with a low false positive rate. Overall design: the tissue-specific transcriptome was analysed by SRT method using U2-3 promoter for multiple-tissue profiling, myo-3 promoter for muscle-specific profiling, rgef-1 promoter for pan-neuron specific profiling with 2 biological replicates for most cases.
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