A5053601159- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Advanced biosensing and bioanalysis techniques
- Insect symbiosis and bacterial influences
- RNA Interference and Gene Delivery
- Innovation and Socioeconomic Development
- Bacteriophages and microbial interactions
- Gene Regulatory Network Analysis
- Microbial Metabolic Engineering and Bioproduction
- Plant Virus Research Studies
- Mosquito-borne diseases and control
- Vibrio bacteria research studies
- RNA regulation and disease
- Viral Infections and Immunology Research
- Invertebrate Immune Response Mechanisms
- Probiotics and Fermented Foods
- Gut microbiota and health
- Transgenic Plants and Applications
- Animal Genetics and Reproduction
- RNA modifications and cancer
- Evolution and Genetic Dynamics
- Insect Utilization and Effects
- Insect behavior and control techniques
- Single-cell and spatial transcriptomics
University of Würzburg
2018-2025
Helmholtz Institute for RNA-based Infection Research
2018-2025
Helmholtz Centre for Infection Research
2019-2025
North Carolina State University
2015-2024
Universitätsklinikum Würzburg
2022
Senckenberg Society for Nature Research
2021
John Wiley & Sons (United States)
2019
Hudson Institute
2019
University of North Carolina at Chapel Hill
2015
Eunice Kennedy Shriver National Institute of Child Health and Human Development
2010-2012
Abstract CRISPR–Cas9 represents a promising platform for genome editing, yet means its safe and efficient delivery remain to be fully realized. A novel vehicle that simultaneously delivers the Cas9 protein single guide RNA (sgRNA) is based on DNA nanoclews, yarn‐like nanoparticles are synthesized by rolling circle amplification. The biologically inspired vehicles were efficiently loaded with Cas9/sgRNA complexes delivered nuclei of human cells, thus enabling targeted gene disruption while...
CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems in bacteria and archaea employ RNAs to specifically recognize the complementary DNA of foreign invaders, leading sequence-specific cleavage or degradation target DNA. Recent work has shown that accidental intentional targeting bacterial genome is cytotoxic can lead cell death. Here, we have demonstrated with CRISPR-Cas be employed for titratable removal individual strains species. Using type...
CRISPR-Cas systems have shown tremendous promise as heterologous tools for genome editing and transcriptional regulation. Because these RNA-directed immune are found in most prokaryotes, an opportunity exists to harness the endogenous convenient organisms. Here, we report that Type I-E system Escherichia coli can be co-opted programmable repression. We deletion of signature cas3 gene converted this into a regulator capable reversible silencing genes. Targeting promoter regions yielded...
Cellular RNAs guide CRISPR-Cas9 The Cas9 nuclease widely used for genome editing is derived from natural bacterial defense systems that protect against invading viruses. directed by RNA guides to cut matching viral DNA. Jiao et al. discovered can also originate cellular unassociated with (see the Perspective Abudayyeh and Gootenberg). They rendered this process programmable, linking presence of virtually any cutting DNA Cas9. This capability basis a new CRISPR diagnostic method developed...
Abstract Bacterial abortive-infection systems limit the spread of foreign invaders by shutting down or killing infected cells before can replicate 1,2 . Several RNA-targeting CRISPR–Cas (that is, types III and VI) cause phenotypes activating indiscriminate nucleases 3–5 However, a CRISPR-mediated abortive mechanism that leverages DNase activity an RNA-guided single-effector nuclease has yet to be observed. Here we report RNA targeting type V Cas12a2 drives infection through non-specific...
Abstract Cas12a2 is a CRISPR-associated nuclease that performs RNA-guided, sequence-nonspecific degradation of single-stranded RNA, DNA and double-stranded following recognition complementary RNA target, culminating in abortive infection 1 . Here we report structures binary, ternary quaternary complexes to reveal complete activation pathway. Our autoinhibited until binding cognate which exposes the RuvC active site within large, positively charged cleft. Double-stranded substrates are...
Abstract CRISPR interference (CRISPRi) is the leading technique to silence gene expression in bacteria; however, design rules remain poorly defined. We develop a best-in-class prediction algorithm for guide silencing efficiency by systematically investigating factors influencing depletion genome-wide essentiality screens, with surprising discovery that gene-specific features substantially impact prediction. mixed-effect random forest regression model provides better estimates of efficiency....
Animal-microbe facultative symbioses play a fundamental role in ecosystem and organismal health. Yet, due to the flexible nature of their association, selection pressures that act on animals symbionts remain elusive. Here we apply experimental evolution Drosophila melanogaster associated with its growth-promoting symbiont Lactobacillus plantarum, representing well-established model symbiosis. We find diet host, rather than host itself, is predominant driving force this Furthermore, identify...
Abstract CRISPR–Cas9 represents a promising platform for genome editing, yet means its safe and efficient delivery remain to be fully realized. A novel vehicle that simultaneously delivers the Cas9 protein single guide RNA (sgRNA) is based on DNA nanoclews, yarn‐like nanoparticles are synthesized by rolling circle amplification. The biologically inspired vehicles were efficiently loaded with Cas9/sgRNA complexes delivered nuclei of human cells, thus enabling targeted gene disruption while...
Lactic-acid bacteria such as Lactobacillus plantarum are commonly used for fermenting foods and probiotics, where increasingly sophisticated genome-editing tools employed to elucidate enhance these microbes' beneficial properties. The most advanced date utilize an oligonucleotide or double-stranded DNA donor recombineering Cas9 targeted cleavage. As the associated methods often developed in isolation one strain, it remains unclear how different Cas9-based editing compare across strains....
Abstract CRISPR-Cas systems inherently multiplex through CRISPR arrays—whether to defend against different invaders or mediate multi-target editing, regulation, imaging, sensing. However, arrays remain difficult generate due their reoccurring repeat sequences. Here, we report a modular, one-pot scheme called CRATES construct and array libraries. allows assembly of repeat-spacer subunits using defined junctions within the trimmed portion spacers. Using CRATES, for single-effector nucleases...
Escherichia coli cell-free transcription-translation (TXTL) systems offer versatile platforms for advanced biomanufacturing and prototyping synthetic biological parts devices. Production testing could be accelerated with the use of linear DNA, which can rapidly cheaply synthesized. However, DNA is efficiently degraded in TXTL preparations from E. coli. Here, we show that double-stranded encoding χ sites-eight base-pair sequences preferentially bound by RecBCD recombination...