A5039848587- CRISPR and Genetic Engineering
- Advanced biosensing and bioanalysis techniques
- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- SARS-CoV-2 detection and testing
- Biosensors and Analytical Detection
- Cancer Genomics and Diagnostics
- Microbial Natural Products and Biosynthesis
- Pluripotent Stem Cells Research
- Plant-Microbe Interactions and Immunity
- Microbial Metabolic Engineering and Bioproduction
- Microbial Metabolism and Applications
- Genetic factors in colorectal cancer
- Legume Nitrogen Fixing Symbiosis
- Innovation and Socioeconomic Development
- Advanced Biosensing Techniques and Applications
- Plant Disease Resistance and Genetics
- Chromosomal and Genetic Variations
- Medical Imaging and Pathology Studies
- RNA modifications and cancer
- Retinal Development and Disorders
- Chemical Synthesis and Analysis
- Marine Sponges and Natural Products
- Genomics and Phylogenetic Studies
- Mosquito-borne diseases and control
University of Illinois Urbana-Champaign
2020-2025
Shanghai Jiao Tong University
2018-2021
Nanjing Agricultural University
2014
Abstract The need for rapid, accurate, and scalable testing systems COVID-19 diagnosis is clear urgent. Here, we report a rapid S calable Po rtable T esting (SPOT) system consisting of highly sensitive, accurate assay battery-powered portable device diagnosis. SPOT comprises one-pot reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) followed by Pf Ago-based target sequence detection. It capable detecting the N gene E in multiplexed reaction with limit detection (LoD) 0.44...
Technological advances in rare DNA mutations detection have revolutionized the diagnosis and monitoring of tumors, but they are still limited by lack supersensitive high-coverage procedures for identifying low-abundance mutations. Here, we describe a single-tube, multiplex PCR-based system, A-Star, that involves hyperthermophilic Argonaute from Pyrococcus furiosus (PfAgo) highly efficient beneficial its compatibility with polymerase. This novel technique uses specific guide design strategy...
Abstract Argonaute proteins (Agos) from thermophiles function as endonucleases via guide-target base-pairing cleavage for host defense. Since guides play a key role in regulating the catalytic specificity of Agos, elucidating its underlying molecular mechanisms would promote application Agos medical sciences. Here, we reveal that an Ago Pyrococcus furiosus ( Pf Ago) showed stepwise endonuclease activity, which was demonstrated through double-stranded DNA directed by single guide (gDNA)...
The CRISPR-Cas12a system is more advantageous than the widely used CRISPR-Cas9 in terms of specificity and multiplexibility. However, its on-target editing efficiency typically much lower that system. Here we improved by simply incorporating 2-aminoadenine (base Z, which alters canonical Watson-Crick base pairing) into crRNA to increase binding affinity between complementary DNA target. resulting (named zCRISPR-Cas12a thereafter) shows an comparable but with off-target effects mammalian...
Abstract The CRISPR/Cas system has emerged as a powerful tool for genome editing in metabolic engineering and human gene therapy. However, locating the optimal site on chromosome to integrate heterologous genes using remains an open question. Selecting suitable integration involves considering multiple complex criteria, including factors related CRISPR/Cas-mediated integration, genetic stability, expression. Consequently, identifying such sites specific or different chromosomal locations...
CRISPR/Cas9 is a powerful genome editing tool, but its off-target cleavage activity can result in unintended adverse outcomes for therapeutic applications. Here we report the design of simple tunable CRISPR controller which chemically inducible anti-CRISPR protein AcrIIA4 engineered to disable Cas9 DNA binding upon addition trimethoprim. Dose-dependent control over and dCas9 induction was achieved, drastically improved specificity biosafety system. We utilized as means interfere with...
Natural products (NPs) produced by bacteria, fungi and plants are a major source of drug leads. Streptomyces species particularly important in this regard as they produce numerous natural with prominent bioactivities. Here we report fully utomated, s calable high-throughput platform for discovery bioactive n atural p roducts S treptomyces (FAST-NPS). This comprises computational prediction prioritization target biosynthetic gene clusters (BGCs) guided self-resistance genes, highly efficient...
Abstract Thermophilic Argonaute proteins (Agos) can function as endonucleases via specific guide-target base-pairing cleavage for host defense. The ability to cleave target DNA sequences at any arbitrary sites endows them with reprogramed capacity. Here, we identify that an Ago from the hyperthermophilic archaeon Pyrococcus furiosus ( Pf Ago) shows a stepwise endonuclease activity, which is demonstrated by double strand directed single guide rather than canonical one pair of DNAs. We reveal...
Abstract The ability to detect rare mutations has revolutionized diagnosis and monitoring of tumors, but is limited by the shortage sensitive, cost-effective high coverage methods for identification extremely low abundant mutations. Here, we establish a single-tube multiplex PCR system employing thermophilic Argonaute-derived DNA-guided nuclease highly efficient mutation detection, referred as A-Star (Argonaute-directed specific target enrichment detection), that combines selective cleavage...
Abstract The CRISPR/Cas system has emerged as a powerful tool for genome editing in metabolic engineering and human gene therapy. However, locating the optimal site on chromosome to integrate heterologous genes using remains an open question. Selecting suitable integration involves considering multiple complex criteria, including factors related CRISPR/Cas-mediated integration, genetic stability, expression. Consequently, identifying such sites specific or different chromosomal locations...
The Argonaute protein from the archaeon Pyrococcus furiosus (PfAgo) is a DNA-guided nuclease that targets DNA with any sequence. We designed virus detection assay in which PfAgo enzyme cleaves reporter probe thus generating fluorescent signals when amplicons reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) contain target sequences. confirmed RT-LAMP-PfAgo for Delta variant produced significantly higher (p < 0.001) single nucleotide polymorphism (SNP), exclusive to...
The need for rapid, accurate, and scalable testing systems COVID-19 diagnosis is clear urgent. Here we report a rapid Scalable Portable Testing (SPOT) system consisting of highly sensitive, accurate assay battery-powered portable device diagnosis. SPOT comprises one-pot reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) followed by PfAgo-based target sequence detection. It capable detecting the N gene E in multiplexed reaction with limit detection (LoD) 0.44 copies/μL...