promoter selection in plasmid based shRNA - (Aug/30/2005 )
I am new here. I hope someone can tell me, if the promoter we choosed to lead the short hairpin RNA is strictly restrict to one kind of promoter,such as U6 and H1. I know this promoter is weak. Is anyone try to use the the CMV promoter? or it can not be used? Hoping for your reply! Thank you!
several papers refers to use of CMV promoter.
Twoapproaches were used. First one CMV enhancer sequence coupled to U6 promoter Xia et al.
Second one, directly full CMV promoter sequence.
Here are few refs :
Xia, X. G., Zhou, H., Ding, H., Affar el, B., Shi, Y., and Xu, Z. (2003). An enhanced U6 promoter for synthesis of short hairpin RNA. Nucleic Acids Res 31, e100.
Wang, J., Tekle, E., Oubrahim, H., Mieyal, J.J., Stadtman, E.R. and Chock, P.B. (2003) Stable and controllable RNA interference: Investigating the physiological function of glutathionylated actin. Proc Natl Acad Sci U S A, 100, 5103-6.
Diallo, M., Arenz, C., Schmitz, K., Sandhoff, K. and Schepers, U. (2003) Long endogenous dsRNAs can induce complete gene silencing in mammalian cells and primary cultures. Oligonucleotides, 13, 381-92.
Xia, H., Mao, Q., Paulson, H.L. and Davidson, B.L. (2002) siRNA-mediated gene silencing in vitro and in vivo. Nat Biotechnol, 20, 1006-10.
Thank you very much! I think you reply is very useful for my experiment! Thank you!