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. 2014 Jan 27;5(2):e27913. doi: 10.4161/trns.27913

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Copyright © 2014 Landes Bioscience

This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.

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graphic file with name tran-5-e27913-g7.jpg — Figure 7. Coligo transcript processing by Dicer. (A) Sequences and predicted secondary structures of two additional coligos used for IVT in the presence of recombinant human Dicer. Shaded regions indicate cDNA of mature miRNA 21 and 143. TSS, predicted transcription start site; LL, larger loop; black triangle, circularization site; nt, coligo sizes in nucleotides. (B) HEK293T WCE IVT in the presence or absence of supplemental recombinant Dicer (rDcr). Bracket indicates gel region containing expected monomer transcripts. M, marker; L, linear precursor; C, coligo. (C) DPAGE analysis of an in vitro processing reaction between coligo 122’s purified monomer transcript and FLAG-Dicer immunoprecipitated from transiently transfected HEK293T cells. (D) Predicted secondary structures of coligo 122 and its monomer transcript, with deduced Dicer cut sites indicated.