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

被引：1
|
作者：
Hueso, Miguel [1 ,2 ]
Mallen, Adrian [3 ]
Ripoll, Elia [3 ]
de Ramon, Laura [3 ]
Bolanos, Nuria [3 ]
Varela, Cristian [3 ]
Guiteras, Jordi [3 ]
Checa, Javier [4 ]
Navarro, Estanislao
Maria Grinyo, Josep [5 ]
Maria Cruzado, Josep [1 ,2 ,5 ]
Maria Aran, Josep [4 ]
Torras, Joan [1 ,2 ,5 ]
机构：
[1] Hosp Univ Bellvitge, Dept Nephrol, Barcelona, Spain
[2] Inst Invest Biomed Bellvitge IDIBELL, Barcelona, Spain
[3] Inst Invest Biomed Bellvitge IDIBELL, Expt Nephrol Lab, Barcelona, Spain
[4] Inst Invest Biomed Bellvitge IDIBELL, Immunoinflammatory Proc & Gene Therapeut Lab, Barcelona, Spain
[5] Univ Barcelona, Dept Clin Sci, Bellvitge Campus, Barcelona, Spain
来源：
BIO-PROTOCOL | 2021年 / 11卷 / 10期
关键词：
siRNA; CD40; CD40L; Cholesterol-derived oligonucleotide; Kidney; siRNA therapy; Off-target effects; MECHANISM;
D O I：
10.21769/BioProtoc.4032
中图分类号：
Q [生物科学];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
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.
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页数：34
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