In vitro selection and characterization of cellulose-binding DNA aptamers.
Many nucleic acid enzymes and aptamers have modular architectures that allow them to retain their functions when combined with other nucleotide sequences. This modular function facilitates the engineering of RNAs and DNAs that have more complex functions. We sought to create new DNA aptamers that bind cellulose to provide a module for immobilizing DNAs. Cellulose has been used in a variety of applications ranging from coatings and films to pharmaceutical preparations, and therefore DNA aptamers that bind cellulose might enable new applications. We used in vitro selection to isolate aptamers from a pool of random-sequence DNAs and subjected two distinct clones to additional rounds of mutagenesis and selection. One aptamer (CELAPT 14) was chosen for sequence minimization and more detailed biochemical analysis. CELAPT 14 aptamer variants exhibit robust binding both to cellulose powder and paper. Also, an allosteric aptamer construct was engineered that exhibits ATP-mediated cellulose binding during paper chromatography.
Boese
Benjamin J
BJ
Department of Chemical Engineering, Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06520-8103, USA.
Breaker
Ronald R
RR
D000494:Allosteric Regulation; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification*; D001483:Base Sequence; D002482:Cellulose / Q000737:chemistry*; D002846:Chromatography, Affinity; D002854:Chromatography, Paper; D004252:DNA Mutational Analysis; D054856:G-Quadruplexes; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation
D052157:Aptamers, Nucleotide; D002482:Cellulose