RNA interference in keratinocytes and an organotypic model of human epidermis.

Cory L. Simpson*, Shin ichiro Kojima, Spiro Getsios

*Corresponding author for this work

Research output: Contribution to journalArticle

33 Scopus citations

Abstract

Gene silencing approaches afford investigators the ability to gain important insight into the normal functional requirements of specific epidermal proteins and promise to yield a powerful therapeutic means to dampen the level of proteins that are mutated or frequently overexpressed in skin disease. The efficient and tractable delivery of siRNAs into epidermal keratinocytes is seminal to this process. Here, we describe techniques for transient and long-term silencing of a representative gene product, namely desmoglein 1, in primary human epidermal keratinocytes maintained as submerged cultures or three-dimensional organotypic raft cultures. As a complement to epidermal-specific gene targeting strategies in mice, these technical approaches permit relatively rapid loss-of-function studies purely in keratinocytes without some of the potential influences present in situ, such as an immune system or vasculature.

Original languageEnglish (US)
Pages (from-to)127-146
Number of pages20
JournalMethods in molecular biology (Clifton, N.J.)
Volume585
StatePublished - Jan 1 2010

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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