Optical antisense tumor targeting in vivo with an improved fluorescent DNA duplex probe
Department of Radiology
Medical Subject Headings
Animals; Base Sequence; Benzothiazoles; Cell Line, Tumor; DNA Probes; DNA, Antisense; Fluorescence; Humans; Male; Mice; Neoplasms; Nucleic Acid Hybridization; Quinolines
Chemistry | Neoplasms | Radiology
Fluorescent conjugated DNA oligonucleotides for antisense targeting of mRNA has the potential of improving tumor/normal tissue ratios over that achievable by nuclear antisense imaging. By conjugating the Cy5.5 emitter to the 3' equivalent end of a 25 mer phosphorothioate (PS) antisense major DNA and hybridizing with a shorter 18 mer phosphodiester (PO) complementary minor DNA (cDNA) with the Black Hole inhibitor BHQ3 on its 5' end (i.e., PS DNA25-Cy5.5/PO cDNA18-BHQ3), we previously achieved antisense optical imaging in mice as a proof of this concept. In a process of optimization, we have now evaluated the stability of a small series of duplexes with variable-length minor strands. From these results, a new study anti-mdr1 antisense duplex was selected with a 10 mer minor strand (i.e., PS DNA25-Cy5.5/PO cDNA10-BHQ3). The new study duplex shows stability in serum environments at 37 degrees C and provides a dramatically enhanced fluorescence in KB-G2 (pgp++) cells when compared with KB-31 (pgp+/-) as evidence of antisense dissociation at its mdr1 mRNA target. The duplex was also administered to KB-G2 tumor bearing mice, and when compared to the duplex used previously, the fluorescence from the tumor thigh was more obvious and the tumor-to-background fluorescence ratio was improved. In conclusion, by a process designed to optimize the duplex for optical antisense tumor targeting, the fluorescence signal was improved both in cells and in tumored mice.
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Citation: Bioconjug Chem. 2009 Jun;20(6):1223-7. doi: 10.1021/bc9000933. Link to article on publisher's site
Liang, Min Min; Liu, Xinrong; Cheng, Dengfeng; Nakamura, Kayoko; Wang, Yi; Dou, Shuping; Liu, Guozheng; Rusckowski, Mary; and Hnatowich, Donald J., "Optical antisense tumor targeting in vivo with an improved fluorescent DNA duplex probe" (2009). Radiology Publications and Presentations. 57.