Transfection of GFP mRNA in dendritic cells and analysis of some factors involved

Fei Zhong, Zhen Yu Zhong, Shuang Liang, Xiu Jin Li

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

AIM: To investigate the transfection of green fluorescent protein (GFP) mRNA in dendritic cells (DC) and analyze some factors which influence the transfection efficiency. METHODS: GFP (as a report gene) mRNA with cap was synthesized, in vitro, with mMESSAGE RNA Transcription Kit containing T7 RNA polymerase, and then the poly(A) was added to the GFP caped-mRNA by yeast poly(A) polymerase. DC were generated from the monocytes isolated from human peripheral blood by stimulation of GM-CSF and IL-4. The GFP mRNA was transfected into DC mediated by transfection reagent. The transfection efficiency and the expression levels were measured by flow cytometry. RESULTS: GFP expression in DC has been obtained by transfecting its mRNA synthesized in vitro. The transfection reagents, mRNA concentrations and cell densities have the significant effects on the transfection. The high level of transfection efficiency (up to 27%) was obtained using Transmessenger Transfection Kit with 1 microg gfp mRNA in 200 microL X-VIVO-15 serum-free medium at the cell density of 2.5x10(9)/L. CONCLUSION: The high-level transfection efficiency of gfp gene in DC could been achieved by using GFP mRNA in the optimum transfection conditions.

Original languageEnglish (US)
Pages (from-to)716-719
Number of pages4
JournalXi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Volume22
Issue number6
StatePublished - Nov 2006
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)

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