RRC ID 55856
Author Alberti C, Manzenreither RA, Sowemimo I, Burkard TR, Wang J, Mahofsky K, Ameres SL, Cochella L.
Title Cell-type specific sequencing of microRNAs from complex animal tissues.
Journal Nat Methods
Abstract MicroRNAs (miRNAs) play an essential role in the post-transcriptional regulation of animal development and physiology. However, in vivo studies aimed at linking miRNA function to the biology of distinct cell types within complex tissues remain challenging, partly because in vivo miRNA-profiling methods lack cellular resolution. We report microRNome by methylation-dependent sequencing (mime-seq), an in vivo enzymatic small-RNA-tagging approach that enables high-throughput sequencing of tissue- and cell-type-specific miRNAs in animals. The method combines cell-type-specific 3'-terminal 2'-O-methylation of animal miRNAs by a genetically encoded, plant-specific methyltransferase (HEN1), with chemoselective small-RNA cloning and high-throughput sequencing. We show that mime-seq uncovers the miRNomes of specific cells within Caenorhabditis elegans and Drosophila at unprecedented specificity and sensitivity, enabling miRNA profiling with single-cell resolution in whole animals. Mime-seq overcomes current challenges in cell-type-specific small-RNA profiling and provides novel entry points for understanding the function of miRNAs in spatially restricted physiological settings.
Volume 15(4)
Pages 283-289
Published 2018-4
DOI 10.1038/nmeth.4610
PII nmeth.4610
PMID 29481550
PMC PMC5886366
MeSH Animals Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism Caenorhabditis elegans / genetics* Cloning, Molecular Drosophila / genetics* MicroRNAs / genetics* Neurons / metabolism Sequence Analysis, RNA / methods*
IF 28.467
Times Cited 18
C.elegans tm4477