A5018259778- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- Bacteriophages and microbial interactions
- DNA and Nucleic Acid Chemistry
- Innovative Microfluidic and Catalytic Techniques Innovation
- Microfluidic and Capillary Electrophoresis Applications
- RNA and protein synthesis mechanisms
- Electrowetting and Microfluidic Technologies
- Molecular Biology Techniques and Applications
- Science, Research, and Medicine
- RNA modifications and cancer
- Enzyme Structure and Function
- Chemistry and Chemical Engineering
University of California, Irvine
2017-2020
Illumina (United States)
2015
Synthetic biology aims to improve human health and the environment by repurposing biological enzymes for use in practical applications. However, natural often function with suboptimal activity when engineered into pathways or challenged recognize unnatural substrates. Overcoming this problem requires efficient directed evolution methods discovering new enzyme variants that a desired activity. Here, we describe construction, validation, application of fluorescence-activated droplet sorting...
Abstract The isolation of synthetic genetic polymers (XNAs) with catalytic activity demonstrates that catalysis is not limited to natural biopolymers, but it remains unknown whether such systems can achieve robust Michaelis-Menten kinetics. Here, we describe an efficient RNA-cleaving 2’-fluoroarabino nucleic acid enzyme (FANAzyme) functions a rate enhancement >10 6 -fold over the uncatalyzed reaction and exhibits substrate saturation kinetics typical most enzymes. FANAzyme was generated...
Darwinian evolution experiments carried out on xeno-nucleic acid (XNA) polymers require engineered polymerases that can faithfully and efficiently copy genetic information back forth between DNA XNA. However, current XNA function with inferior activity relative to their natural counterparts. Here, we report five X-ray crystal structures illustrate the pathway by which α-(L)-threofuranosyl nucleic (TNA) triphosphates are selected extended in a template-dependent manner using...
Engineering polymerases to synthesize artificial genetic polymers with unique backbone structures is limited by a general lack of understanding about the structural determinants that govern substrate specificity. Here, we report high-throughput microfluidic-based approach for mapping sequence–function relationships combines droplet-based optical polymerase sorting deep mutational scanning. We applied this strategy map finger subdomain replicative DNA isolated from Thermococcus kodakarensis...
Most DNA polymerase libraries sample unknown portions of mutational space and are constrained by the limitations random mutagenesis. Here we describe a programmed allelic mutagenesis (PAM) strategy to comprehensively evaluate all possible single-point mutations in entire catalytic domain replicative polymerase. By applying PAM with ultrafast high-throughput screening, show how polymerases can be mapped for that exhibit enhanced activity unnatural nucleic acid substrates. We suggest...
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...
Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing. However, whole-genome is still costly and complex for diagnostics purposes. In clinical space, targeted advantage allowing researchers to focus on specific genes interest. Routine use NGS mandates inexpensive instruments, fast turnaround time an integrated robust workflow. Here we demonstrate a version Sequencing Synthesis (SBS) chemistry that potentially can become preferred method in...