Aptamer-derived nucleic acid oligos: applications to develop nucleic acid chips to analyze proteins and small ligands.
The specificity and affinity of aptamers for their cognate ligands are comparable to those of antibodies for antigens. To use aptamers effectively in high-throughput assays in a microarray format, to analyze various analytes, we developed a strategy in which the aptamer was split into two nonfunctional units and allowed to reassemble into the functional aptamer by the cognate ligand. We have named this method "analyte-dependent oligonucleotide modulation assay" (ADONMA). As proof-of-principle, we used oligonucleotides derived from the aptamer RNA against HIV-1 Tat and demonstrated, with both titer plates and plastic slide chips, that specifically in the presence of Tat or its peptide, the two oligos reconstituted the core binding regions of Tat. Thus, these results suggest that ADNOMA has the potential for use in nucleic acid microarrays for detecting various ligands.
Yamamoto-Fujita
Rika
R
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1 Higashi, Tsukuba Science City 305-8566, Japan.
Kumar
Penmetcha K R
PK
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D015696:Gene Products, tat / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D008024:Ligands; D046208:Microchip Analytical Procedures / Q000379:methods*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D017186:Titrimetry
D052157:Aptamers, Nucleotide; D015696:Gene Products, tat; D008024:Ligands; D009696:Nucleic Acids; D009841:Oligonucleotides; D011506:Proteins