In vivo CD40 Silencing by siRNA Infusion in Rodents and Evaluation by Kidney Immunostaining
Miguel Hueso,
Adrian Mallen,
Elia Ripoll,
Laura de Ramon,
Nuria Bolaños,
Christian Valera,
Jordi Guiteras,
Javier Raya,
Estanislao Navarro,
Josep Maria Grinyo,
Josep Maria Cruzado,
Josep Maria Aran,
JUAN TORRAS
Affiliations
Miguel Hueso
Department of Nephrology, Hospital Universitari Bellvitge, and Institut d’Investigació Biomèdica de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Adrian Mallen
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Elia Ripoll
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Laura de Ramon
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Nuria Bolaños
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Christian Valera
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Jordi Guiteras
Experimental Nephrology Laboratory, Institut d’Investigació Biomèdicas de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Javier Raya
Immunoinflammatory Processes and Gene Therapeutics Lab, Institut d’Investigació Biomèdica de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
Estanislao Navarro
Independent researcher, Barcelona, Spain
Josep Maria Grinyo
University of Barcelona, Department of Clinical Sciences, Bellvitge Campus, L’Hospitalet de Llobregat, Barcelona, Spain
Josep Maria Cruzado
Department of Nephrology, Hospital Universitari Bellvitge, and Institut d’Investigació Biomèdica de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, SpainUniversity of Barcelona, Department of Clinical Sciences, Bellvitge Campus, L’Hospitalet de Llobregat, Barcelona, Spain
Josep Maria Aran
Immunoinflammatory Processes and Gene Therapeutics Lab, Institut d’Investigació Biomèdica de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, Spain
JUAN TORRAS
Department of Nephrology, Hospital Universitari Bellvitge, and Institut d’Investigació Biomèdica de Bellvitge-IDIBELL, L’Hospitalet de Llobregat, Barcelona, SpainUniversity of Barcelona, Department of Clinical Sciences, Bellvitge Campus, L’Hospitalet de Llobregat, Barcelona, Spain
The co-stimulatory molecule CD40 and its ligand CD40L play a key role in the regulation of immunological processes and are involved in the pathophysiology of autoimmune and inflammatory diseases. Inhibition of the CD40-CD40L axis is a promising therapy, and a number of strategies and techniques have been designed to hinder its functionality. Our group has broad experience in silencing CD40 using RNAi technology, and here we summarize protocols for the systemic administration of a specific anti-CD40 siRNA in different rodents models, in addition to the subsequent quantification of CD40 expression in murine kidneys by immunostaining. The use of RNAi technology with specific siRNAs to silence genes is becoming an essential method to investigate gene functions and is rapidly emerging as a therapeutic tool.Graphic abstract:CD40 siRNA mechanism
