In vitro selection and characterization of TCF-1 binding RNA aptamers.
TCF proteins are architectural transcription factors that bind to beta-catenin when the Wnt signaling pathway is activated. TCF-1 is specifically expressed in T-cell lineages and its interaction with beta-catenin is thought to be critical for early T-cell development. However, the mechanisms underlying activation of TCF-1 during T-cell development are not completely understood. With the aim of developing RNA aptamers that bind to TCF-1 and regulate its activity, we screened an RNA library consisting of random sequences of 70 nucleotides and were able to isolate aptamers that bind to TCF-1 with high affinity and specificity. We employed RNase footprinting to characterize the RNA structures and map their binding sites for TCF-1. It is hoped that the selected aptamers will regulate TCF-1 activity in vivo, thereby providing a unique tool for modulating TCF-1 function in early T-cell development.
Lee
Seung-Yeon
SY
Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Seoul 140-714, Korea.
Jeong
Sunjoo
S
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015723:Gene Library; D051821:Lymphoid Enhancer-Binding Factor 1; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D051822:T Cell Transcription Factor 1; D013601:T-Lymphocytes / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D014158:Transcription, Genetic; D051176:beta Catenin
D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D051821:Lymphoid Enhancer-Binding Factor 1; D051822:T Cell Transcription Factor 1; D015534:Trans-Activators; D014157:Transcription Factors; D051176:beta Catenin; D012313:RNA; D004247:DNA; D012260:Ribonucleases