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Gene silencing in a human organotypic skin model.

Abstract：

:Here we present a simple and highly reproducible method which allows the study of the effects of a single gene knockdown in an organotypic skin model. Human keratinocytes (KC) were transfected with backbone-modified short interfering RNAs (siRNAs) specific for vascular endothelial growth factor (VEGF) and matriptase-1. Twenty-four hours later the transfected cells were seeded onto fibroblast collagen suspensions and allowed to build up a multilayered epidermis by culture at the air/medium interface for 7 days. Protein expression of both targeted genes remained down-regulated by more than 80% up to 8 days after transfection. As expected, VEGF knockdown by siRNA did not alter epidermis formation in our organotypic skin model. By contrast ablation of matriptase-1 led to aberrant KC differentiation and impaired filaggrin processing and resulted in an epidermal phenotype closely resembling that of matriptase-1 deficient mouse skin. Our results suggest that siRNA-mediated gene silencing is highly efficient in an organotypic skin model and readily allows the assessment of the roles of individual genes during terminal KC differentiation.

journal_name

Biochem Biophys Res Commun

journal_title

Biochemical and biophysical research communications

authors

Mildner M,Ballaun C,Stichenwirth M,Bauer R,Gmeiner R,Buchberger M,Mlitz V,Tschachler E

doi

10.1016/j.bbrc.2006.07.035

subject

Has Abstract

pub_date

2006-09-15 00:00:00

pages

76-82

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(06)01526-9

journal_volume

348

pub_type

杂志文章

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