A5075273861- Advanced biosensing and bioanalysis techniques
- DNA and Nucleic Acid Chemistry
- Bacteriophages and microbial interactions
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- RNA Interference and Gene Delivery
- Electrowetting and Microfluidic Technologies
- Innovative Microfluidic and Catalytic Techniques Innovation
- Molecular Junctions and Nanostructures
- Microfluidic and Capillary Electrophoresis Applications
- Bacterial Genetics and Biotechnology
- Amino Acid Enzymes and Metabolism
- Origins and Evolution of Life
University of California, Irvine
2015-2020
Arizona State University
2013-2016
Abstract Polymerases that synthesize artificial genetic polymers hold great promise for advancing future applications in synthetic biology. However, engineering natural polymerases to replicate unnatural is a challenging problem. Here we present droplet-based optical polymerase sorting (DrOPS) as general strategy expanding function employs an sensor monitor activity inside the microenvironment of uniform compartment generated by microfluidics. We validated this approach performing complete...
Aptamers are often prone to nuclease digestion, which limits their utility in many biomedical applications. Here we describe a xeno-nucleic acid system based on α-l-threofuranosyl nucleic (TNA) that is completely refractory digestion. The use of an engineered TNA polymerase permitted the isolation functional aptamers bind HIV reverse transcriptase (HIV RT) with KD's ∼0.4-4.0 nM. were identified using display strategy provides powerful genotype-phenotype linkage. remain active presence and...
The emerging field of synthetic genetics provides an opportunity to explore the structural and functional properties genetic polymers by in vitro selection. Limiting this process, however, is availability enzymes that allow for synthesis propagation information present unnatural nucleic acid sequences. Here, we report development a transcription reverse-transcription system can replicate composed threose acids (TNA). TNA potential progenitor RNA which natural ribose sugar found has been...
The ability to synthesize and propagate genetic information encoded in the framework of xeno-nucleic acid (XNA) polymers would inform a wide range topics from origins life synthetic biology. While directed evolution has produced examples engineered polymerases that can accept XNA substrates, these enzymes function with reduced activity relative their natural counterparts. Here, we describe biochemical strategy enables discovery improved for given unnatural polymerase function. Our approach...
Recent advances in polymerase engineering have enabled the replication of xenonucleic acid (XNA) polymers with backbone structures distinct from those found nature. By introducing a selective amplification step into cycle, functional XNA molecules been isolated by vitro selection binding and catalytic activity. Despite these successes, coding decoding genetic information remains limited fidelity efficiency engineered polymerases. In particular, process reverse transcribing back DNA for PCR...
Threose nucleic acid (TNA) is an unnatural genetic polymer capable of undergoing Darwinian evolution to generate folded molecules with ligand-binding activity. This property, coupled a nuclease-resistant backbone, makes TNA attractive candidate for future applications in biotechnology. Previously, we have shown that engineered form the Archaean replicative DNA polymerase 9°N, known commercially as Therminator polymerase, can copy three-letter alphabet (A,T,C) from into TNA. However, our...
Abstract Polymerase engineering is making it possible to synthesize xeno‐nucleic acid polymers (XNAs) with diverse backbone structures and chemical functionality. The ability copy genetic information back forth between DNA XNA has led a new field of science known as synthetic genetics, which aims study the concepts heredity evolution in artificial polymers. Since many polymerases needed are not available commercially, researchers must express purify these enzymes recombinant proteins from E....
Engineered polymerases that can copy genetic information between DNA and xeno-nucleic acids (XNA) hold tremendous value as reagents in future biotechnology applications. However, current XNA function with inferior activity relative to their natural counterparts, indicating polymerase engineering efforts would benefit from new benchmarking assays. Here, we describe a highly parallel, low-cost method for measuring the average rate substrate specificity of standard qPCR instrument. Our...