In vivo CD40 Silencing by siRNA Infusion in Rodents and Evaluation by Kidney Immunostaining

啮齿类动物体内通过 siRNA 输注实现 CD40 沉默并通过肾脏免疫染色进行评估

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作者:Miguel Hueso, Adrián Mallén, Elia Ripoll, Laura de Ramón, Núria Bolaños, Cristian Varela, Jordi Guiteras, Javier Checa, Estanislao Navarro, Josep Maria Grinyo, Josep Maria Cruzado, Josep Maria Aran, Joan Torras

Abstract

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.

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