I’m trying to express some small RNAs in the germline and have been unsuccessfully trying to find a heterologous PolIII regulated promoter for this purpose. Essentially something similar to the mammalian U1, U6, or H promoters. I’ve tried to look into the C. elegans U1 and U6 genes, but they don’t seem to be very well characterized. Does anyone have any suggestions?
We have used the U2-3 promoter to express SL RNAs (about 100 nt). The details are in the following papers:
Harrison, N., Kalbfleisch, A., Connolly, B., Pettitt, J., & Müller, B. (2010). SL2-like spliced leader RNAs in the basal nematode Prionchulus punctatus: New insight into the evolution of nematode SL2 RNAs. RNA (New York, NY), 16(8), 1500–1507. doi:10.1261/rna.2155010
Ferguson, K. C., & Rothman, J. H. (1999). Alterations in the conserved SL1 trans-spliced leader of Caenorhabditis elegans demonstrate flexibility in length and sequence requirements in vivo. Molecular and Cellular Biology, 19(3), 1892–1900.