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10.1101/2020.09.17.302232

http://scihub22266oqcxt.onion/10.1101/2020.09.17.302232
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32995793!7523121!32995793
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suck abstract from ncbi


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pmid32995793      bioRxiv 2021 ; ä (ä): ä
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  • Fixed single-cell RNA sequencing for understanding virus infection and host response #MMPMID32995793
  • Van Phan H; van Gent M; Drayman N; Basu A; Gack MU; Tay S
  • bioRxiv 2021[Jan]; ä (ä): ä PMID32995793show ga
  • Single-cell transcriptomic studies that require intracellular protein staining, rare cell sorting, or inactivation of infectious pathogens are severely limited because current high-throughput RNA sequencing methods are incompatible with paraformaldehyde treatment, a common tissue and cell fixation and preservation technique. Here we present FD-seq, a high-throughput method for droplet-based RNA sequencing of paraformaldehyde-fixed, stained and sorted single-cells. We show that FD-seq preserves the mRNA integrity and relative abundances during fixation and subsequent cell retrieval. Furthermore, FD-seq detects a higher number of genes and transcripts than methanol fixation. We applied FD-seq to investigate two important questions in Virology. First, by analyzing a rare population of cells supporting lytic reactivation of the human tumor virus KSHV, we identified TMEM119 as a host factor that mediates viral reactivation. Second, we found that upon infection with the betacoronavirus OC43, which causes the common cold and is a close relative of SARS-CoV-2, pro-inflammatory pathways are primarily upregulated in lowly-infected cells that are exposed to the virus but fail to express high levels of viral genes. FD-seq thus enables integrating phenotypic with transcriptomic information in rare cell populations, and preserving and inactivating pathogenic samples that cannot be handled under regular biosafety measures.
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