RNA interference in human foreskin fibroblasts within the three-dimensional collagen matrix

Mark A. Carlson, Amy K. Prall, Jeremiah J. Gums

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

The technique of RNA interference (RNAi) was trialed in primary human foreskin fibroblasts, both in monolayer culture and in the fibroblast-populated collagen matrix. Knockdown of lamin A/C, p53, and FAK was possible with low-confluency (<50%) monolayer fibroblasts, a transfection vehicle concentration of 1%, and an siRNA concentration of 25-50 nM. Knockdown also was possible in the collagen matrix using similar reagent concentrations and a cellular density of one million fibroblasts per ml of matrix. Optimization of transfection conditions appeared to be important to increase knockdown efficiency. Consistent with prediction, knockdown of FAK induced apoptosis in the fibroblast-populated collagen matrix.

Original languageEnglish (US)
Pages (from-to)123-132
Number of pages10
JournalMolecular and cellular biochemistry
Volume306
Issue number1-2
DOIs
StatePublished - Dec 2007

Keywords

  • Collagen
  • Extracellular matrix
  • Fibroblast
  • Focal adhesion kinase
  • RNA interference
  • p53

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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