Construction and verification of stable down-regulated MMP9,S100A10 mouse brain microvascular endothelial cell lines

Abstract

Abstract:

Objective To construct stable down-regulated MMP9,S100A10 mouse brain microvascular endothelial cell lines,then verify the reduction level of gene expression.Methods By transfecting LV-musMMP9-shRNA,LV-musS100A10-shRNA into b.End3 respectively,then the very cell lines were screened by puromycin.The obtained differential b.End3 cell lines were observed by fluorescence microscope and varified by RT-qPCR.Results The study successfully constructed stable transfected cell lines which could effectively interference S100A10,MMP-9 expression of 3×10⁸ TU/mL and 1×10⁸ TU/mL LV-musS100A10-shRNA respectively.Conclusions The targeted down-regulation b.End3 cell lines are useful for further exploring the role of molecule S100A10 or MMP-9 in host blood-brain barrier by control variables and the possible target of treatment exploration of cryptococcal encephalitis/meningitis.

Key words: Lentivirus vector, MMP9, S100A10, b.End3

CLC Number:

R392.1

CHEN Xue-wen, ZHU Hong-mei, WEN Hai. Construction and verification of stable down-regulated MMP9,S100A10 mouse brain microvascular endothelial cell lines[J]. Chinese Journal of Mycology, 2017, 12(1): 19-22,18.

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