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NanoSINC-seq dissects the isoform diversity in subcellular compartments of single cells


Abstract
Alternative mRNA isoforms play a key role in generating diverse protein isoforms. To dissect isoform usage in the subcellular compartments of single cells, we introduced an novel approach, nanopore sequencing coupled with single-cell integrated nuclear and cytoplasmic RNA sequencing, that couples microfluidic fractionation, which separates cytoplasmic RNA from nuclear RNA, with full-length complementary DNA (cDNA) sequencing using a nanopore sequencer. Leveraging full-length cDNA reads, we found that the nuclear transcripts are notably more diverse than cytoplasmic transcripts. Our findings also indicated that transcriptional noise emanating from the nucleus is regulated across the nuclear membrane and then either attenuated or amplified in the cytoplasm depending on the function involved. Overall, our results provide the landscape that shows how the transcriptional noise arising from the nucleus propagates to the cytoplasm. ....

Grand Island , United States , France General , Takara Bio , Panreac Applichem , Kyoto University , Marubun Research Promotion Foundation Japan , Exchange Research , Cabinet Office , University Of Tokyo , White Rock Foundation , Wako Pure Chemical Industries , Japan Society For The Promotion Of Science , Oxford Nanopore Technologies , Thermo Fisher Scientific , Dojindo Laboratories , Low Input , Preparation Kit , Ligation Sequencing Kit , Creative Commons Attribution Noncommercial , Score Program , Japan Society , Marubun Research Promotion Foundation , மாபெரும் தீவு , ஒன்றுபட்டது மாநிலங்களில் , பிரான்ஸ் ஜநரல் ,