Intracellular expression of the T-cell factor-1 RNA aptamer as an intramer.
T-cell factor (TCF)-1 protein forms the transcriptional complex with beta-catenin and regulates the expression of diverse target genes during early development and carcinogenesis. We have selected previously an RNA aptamer that binds to the DNA-binding domain of TCF-1 and have shown that it interfered with binding of TCF-1 to its specific DNA recognition sequences in vitro. As an approach to modulate the transcription by TCF/beta-catenin complex in the cells, we have developed the RNA expression vector for stable expression of RNA aptamer inside of the mammalian cells. High level of RNA was expressed as an intramer in the fusion with the stable RNA transcript. The RNA intramer inhibited TCF/beta-catenin transcription activity as shown by luciferase assay. It also modulated the expression of TCF/beta-catenin target genes, such as cyclin D1 and matrix metalloproteinase-7, as predicted to be as an effective inhibitor of the TCF function. In addition, it efficiently reduced the growth rate and tumorigenic potential of HCT116 colon cancer cells. Such RNA intramer could lead to valuable gene therapeutics for TCF/beta-catenin-mediated carcinogenesis.
Choi
Kang Hyun
KH
Department of Molecular Biology, BK21 Graduate Program for RNA Biology, Institute of Nanosensor and Biotechnology, Dankook University, Hannam-dong san 8, Yongsan-ku, Seoul 140-714, Korea.
Park
Min Woo
MW
Lee
Seung Yeon
SY
Jeon
Mi-Ya
MY
Kim
Mee Young
MY
Lee
Hee Kyu
HK
Yu
Jaehoon
J
Kim
Hong-Jin
HJ
Han
Kyungsook
K
Lee
Heviran
H
Park
Keerang
K
Park
Woong June
WJ
Jeong
Sunjoo
S
D052157:Aptamers, Nucleotide / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D048708:Cell Growth Processes / Q000235:genetics; D015870:Gene Expression; D045325:HCT116 Cells; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D051822:T Cell Transcription Factor 1 / Q000096:biosynthesis / Q000235:genetics*; D014158:Transcription, Genetic; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
D052157:Aptamers, Nucleotide; D051822:T Cell Transcription Factor 1; D051176:beta Catenin