siRNA Electroporation to Modulate Autophagy in Herpes Simplex Virus Type 1-Infected Monocyte-Derived Dendritic Cells

Düthorn A, Turan A, Drassner C, Muehl-Zurbes P, Heilingloh C, Steinkasserer A, Grosche L (2019)

Publication Type: Journal article

Publication year: 2019

Journal

Journal of Visualized Experiments MYJoVE Corporation

Journal Issue: 152

Abstract

Herpes simplex virus type-1 (HSV-1) induces autophagy in both, immature dendritic cells (iDCs) as well as mature dendritic cells (mDCs), whereas autophagic flux is only observed in iDCs. To gain mechanistic insights, we developed efficient strategies to interfere with HSV-1-induced autophagic turnover. An inhibitor-based strategy, to modulate HSV-1-induced autophagy, constitutes the first choice, since it is an easy and fast method. To circumvent potential unspecific off-target effects of such compounds, we developed an alternative siRNA-based strategy, to modulate autophagic turnover in iDCs upon HSV-1 infection. Indeed, electroporation of iDCs with FIP200-specific siRNA prior to HSV-1 infection is a very specific and successful method to ablate FIP200 protein expression and thereby to inhibit autophagic flux. Both presented methods result in the efficient inhibition of HSV-1-induced autophagic turnover in iDCs, whereby the siRNA-based technique is more target specific. An additional siRNA-based approach was developed to selectively silence the protein expression of KIF1 B and KIF2A, facilitating autophagic turnover upon HSV-1 infection in mDCs. In conclusion, the technique of siRNA electroporation represents a promising strategy, to selectively ablate the expression of distinct proteins and to analyze their influence upon an HSV-1 infection.

Authors with CRIS profile

Alexandra Birzer Department of Immune Modulation Christiane Heilingloh Lehrstuhl für Biotechnik (Proteinstruktur und -design) Alexander Steinkasserer Professur für Experimentelle Dermatologie

How to cite

APA:

Düthorn, A., Turan, A., Drassner, C., Muehl-Zurbes, P., Heilingloh, C., Steinkasserer, A., & Grosche, L. (2019). siRNA Electroporation to Modulate Autophagy in Herpes Simplex Virus Type 1-Infected Monocyte-Derived Dendritic Cells. Journal of Visualized Experiments, 152. https://doi.org/10.3791/60190

MLA:

Düthorn, Alexandra, et al. "siRNA Electroporation to Modulate Autophagy in Herpes Simplex Virus Type 1-Infected Monocyte-Derived Dendritic Cells." Journal of Visualized Experiments 152 (2019).

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