Deoxyribozymes that recode sequence information.
Allosteric nucleic acid ligases have been used previously to transform analyte-binding into the formation of oligonucleotide templates that can be amplified and detected. We have engineered binary deoxyribozyme ligases whose two components are brought together by bridging oligonucleotide effectors. The engineered ligases can 'read' one sequence and then 'write' (by ligation) a separate, distinct sequence, which can in turn be uniquely amplified. The binary deoxyribozymes show great specificity, can discriminate against a small number of mutations in the effector, and can read and recode DNA information with high fidelity even in the presence of excess obscuring genomic DNA. In addition, the binary deoxyribozymes can read non-natural nucleotides and write natural sequence information. The binary deoxyribozyme ligases could potentially be used in a variety of applications, including the detection of single nucleotide polymorphisms in genomic DNA or the identification of short nucleic acids such as microRNAs.
Tabor
Jeffrey J
JJ
Center for Systems and Synthetic Biology and Institute for Cell and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA.
Levy
Matthew
M
Ellington
Andrew D
AD
D000494:Allosteric Regulation; D020029:Base Pairing; D001483:Base Sequence; D002384:Catalysis; D020134:Catalytic Domain; D011088:DNA Ligases / Q000737:chemistry* / Q000378:metabolism; D021881:DNA, Catalytic / Q000737:chemistry* / Q000378:metabolism; D004789:Enzyme Activation; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015202:Protein Engineering; D013379:Substrate Specificity
D021881:DNA, Catalytic; D009841:Oligonucleotides; D011088:DNA Ligases