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. Author manuscript; available in PMC: 2012 Jun 1.

Published in final edited form as: Methods. 2011 Jan 20;54(2):284–294. doi: 10.1016/j.ymeth.2010.12.039

The design and evaluation of Ab′ pRNA-siRNA chimeras. (A) Schematic of the Ab′ pRNA-siRNA chimera. The siRNA part of the chimera consists of 27 bps. As an example here, the siRNA targets HIV-1 tat/Rev. The chimeric pRNA-siRNA sense strand was transcribed in vitro with T7 RNA polymerase and then annealed with the antisense strand to get the whole pRNA-siRNA chimera. A linker (UU) between the aptamer and siRNA is indicated. (B) The stability assay in cell culture medium containing 10% FBS of the 2′-F modified pRNA-siRNA chimeras vs. unmodified pRNA-siRNA chimeras. (C) Gene silencing activity and strand selectivity of pRNA-siRNA chimeras. Dual luciferase assays of psiCHECK sense and anti-sense targets are shown. All RNAs were normalized to the value of the corresponding buffer control. All the data were represent the averages of triplicate assays.

The design and evaluation of Ab′ pRNA-siRNA chimeras. (A) Schematic of the Ab′ pRNA-siRNA chimera. The siRNA part of the chimera consists of 27 bps. As an example here, the siRNA targets HIV-1 tat/Rev. The chimeric pRNA-siRNA sense strand was transcribed in vitro with T7 RNA polymerase and then annealed with the antisense strand to get the whole pRNA-siRNA chimera. A linker (UU) between the aptamer and siRNA is indicated. (B) The stability assay in cell culture medium containing 10% FBS of the 2′-F modified pRNA-siRNA chimeras vs. unmodified pRNA-siRNA chimeras. (C) Gene silencing activity and strand selectivity of pRNA-siRNA chimeras. Dual luciferase assays of psiCHECK sense and anti-sense targets are shown. All RNAs were normalized to the value of the corresponding buffer control. All the data were represent the averages of triplicate assays.