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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http://dx.doi.org/10.21769/BioProtoc.4032 | DOI Listing |
Cell Tissue Res
January 2025
College of Life Science, Henan Normal University, No. 46, Jianshe Road, Xinxiang 453007, Henan, China.
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Dongguan Key Laboratory of Stem Cell and Regenerative Tissue Engineering, the First Dongguan Affiliated Hospital, School of Basic Medical Sciences, Guangdong Medical University, Dongguan, China.
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West Guangxi Key Laboratory for Prevention and Treatment of High-incidence Diseases, Youjiang Medical University for Nationalities, Baise, Guangxi 533000, China. Electronic address:
Argonaute 2 (Ago2) is a crucial enzyme in the RNA interference (RNAi) pathway, essential for gene silencing via the cleavage of target messenger RNA (mRNA) mediated by microRNA (miRNA) or small interfering RNA (siRNA). The activity of Ago2 is a significant biomarker for various diseases, including cancer and viral infections, necessitating precise monitoring techniques. Traditional methods for detecting Ago2 activity are often cumbersome and lack the necessary sensitivity and specificity for low-abundance targets in complex samples.
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