Protocol

Molecular Dermatology

Volume 961 of the series Methods in Molecular Biology pp 351-361

Date:

RNAi-Mediated Gene Function Analysis in Skin

  • Slobodan BeronjaAffiliated withLaboratory of Mammalian Cell Biology & Development, Howard Hughes Medical Institute, The Rockefeller University
  • , Elaine FuchsAffiliated withLaboratory of Mammalian Cell Biology & Development, Howard Hughes Medical Institute, The Rockefeller University Email author 

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Abstract

We have recently developed a method for RNAi-mediated gene function analysis in skin (Beronja et al., Nat Med 16:821–827, 2010). It employs ultrasound-guided in utero microinjections of lentivirus into the amniotic cavity of embryonic day 9 mice, which result in rapid, efficient, and stable transduction into mouse skin. Our technique greatly extends the available molecular and genetic toolbox for comprehensive functional examination of outstanding problems in epidermal biology. In its simplest form, as a single-gene function analysis via shRNA-mediated gene knockdown, our technique requires no animal mating and may need as little as only a few days between manipulation and phenotypic analysis.

Key words

Lentivirus Intraamniotic In utero In vivo RNAi shRNA RNA interference Ultrasound-guided microinjection