We are planning to do siRNA transfection of 2-3mm aorta and mesenteric artery segment taken from rats to silence STIM1, and ORAI1 gene. This vessel segment will be used in functional study of vascular contractility and myogenic tone using myograph. In literature, people use reverse permeabilization transfection, however, we do not have any experience in transfection and we are not sure about the efficiency of this method especially with multilayer and thick vessels like aorta. I would like to get your help in choosing the right transfection protocol and reagent.
Below are some references using reverse permeabilization transfection method.
1. Raina H, Ella SR & Hill MA (2008).Decreased activity of the smooth muscle Na+/Ca2+ exchanger impairs arteriolar myogenic reactivity. Physiol 586, 1669–1681.
2. Yan Yang Yoshiro Sohma Zahra Nourian Srikanth R. Ella Min Li Aaron Stupica Ronald J. Korthuis Michael J. Davis Andrew P. Braun Michael A. Hill. Mechanisms underlying regional differences in the Ca2+ sensitivity of BKCa current in arteriolar smooth muscle. J Physiol 591.5 (2013) pp 1277–1293.
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