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Mime-seq 2.0: a method to sequence microRNAs from specific mouse cell types.

Ariane MandlbauerQiong SunNiko PopitschTanja A SchwickertMiroslava SpanovaJingkui WangStefan L AmeresMeinrad BusslingerLuisa Cochella
Published in: The EMBO journal (2024)
Many microRNAs (miRNAs) are expressed with high spatiotemporal specificity during organismal development, with some being limited to rare cell types, often embedded in complex tissues. Yet, most miRNA profiling efforts remain at the tissue and organ levels. To overcome challenges in accessing the microRNomes from tissue-embedded cells, we had previously developed mime-seq (miRNome by methylation-dependent sequencing), a technique in which cell-specific miRNA methylation in C. elegans and Drosophila enabled chemo-selective sequencing without the need for cell sorting or biochemical purification. Here, we present mime-seq 2.0 for profiling miRNAs from specific mouse cell types. We engineered a chimeric RNA methyltransferase that is tethered to Argonaute protein and efficiently methylates miRNAs at their 3'-terminal 2'-OH in mouse and human cell lines. We also generated a transgenic mouse for conditional expression of this methyltransferase, which can be used to direct methylation of miRNAs in a cell type of choice. We validated the use of this mouse model by profiling miRNAs from B cells and bone marrow plasma cells.
Keyphrases
  • single cell
  • rna seq
  • cell therapy
  • genome wide
  • mouse model
  • bone marrow
  • induced apoptosis
  • dna methylation
  • stem cells
  • squamous cell carcinoma
  • poor prognosis
  • drug delivery
  • signaling pathway
  • cell death
  • binding protein