Visualization of global RNA synthesis in a human (mini-) organ in situ by click chemistry

Talveen S. Purba, Jack Marsh, Kayumba Ng'andu, Svitlana Kurinna, Ralf Paus

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

RNA synthesis can be detected by 5-ethynyl uridine (EU) incorporation and click chemistry. Despite identifying a fundamental functional process, this technique has yet to be widely applied to complex human tissue systems. By incorporating EU into human hair follicle (HF) organs cultured ex vivo, nascent RNA synthesis was detected in situ. EU differentially incorporated across the HF epithelium. Interestingly, RNA synthesis did not correlate with protein synthesis, proliferation or epithelial progenitor cell marker expression. By treating human HFs with the cytotoxic cell cycle inhibitor (R)-CR8, which inhibits transcriptional regulators CDK7 and CDK9, it was further shown that this technique can be used to sensitively detect changes in global RNA synthesis in situ. Together, this work delineates new insights into nascent RNA synthesis within a human (mini)- organ and describes a novel read-out parameter that will enrich future ex vivo human tissue research studies.

Original languageEnglish (US)
Pages (from-to)97-100
Number of pages4
JournalBioTechniques
Volume65
Issue number2
DOIs
StatePublished - Aug 1 2018
Externally publishedYes

Fingerprint

Click Chemistry
Visualization
RNA
Uridine
Hair Follicle
Tissue
Cell Cycle
Stem Cells
Epithelium
Epithelial Cells
Cells
Research
Proteins

Keywords

  • 5-ethynyl uridine
  • click chemistry
  • human hair follicle
  • O-propargyl-puromycin
  • protein synthesis
  • RNA synthesis

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Visualization of global RNA synthesis in a human (mini-) organ in situ by click chemistry. / Purba, Talveen S.; Marsh, Jack; Ng'andu, Kayumba; Kurinna, Svitlana; Paus, Ralf.

In: BioTechniques, Vol. 65, No. 2, 01.08.2018, p. 97-100.

Research output: Contribution to journalArticle

Purba, Talveen S. ; Marsh, Jack ; Ng'andu, Kayumba ; Kurinna, Svitlana ; Paus, Ralf. / Visualization of global RNA synthesis in a human (mini-) organ in situ by click chemistry. In: BioTechniques. 2018 ; Vol. 65, No. 2. pp. 97-100.
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