27 results on '"Citorik, Robert"'
Search Results
2. Stable transduction of the neonatal mouse liver using a hybrid rAAV/sleeping beauty transposon gene delivery system.
- Author
-
Cunningham, Sharon C., Zakas, Philip M., Sasaki, Natsuki, van Dijk, Eva B., Zhu, Erhua, Fu, Yanfang, Salomon, William E., Citorik, Robert J., Rubens, Jacob R., Cotta‐Ramusino, Cecilia, Querbes, William, and Alexander, Ian E.
- Abstract
Background: Conventional adeno‐associated viral (AAV) vectors, while highly effective in quiescent cells such as hepatocytes in the adult liver, confer less durable transgene expression in proliferating cells owing to episome loss. Sustained therapeutic success is therefore less likely in liver disorders requiring early intervention. We have previously developed a hybrid, dual virion approach, recombinant AAV (rAAV)/piggyBac transposon system capable of achieving stable gene transfer in proliferating hepatocytes at levels many fold above conventional AAV vectors. An alternative transposon system, Sleeping Beauty, has been widely used for ex vivo gene delivery; however liver‐targeted delivery using a hybrid rAAV/Sleeping Beauty approach remains relatively unexplored. Methods: We investigated the capacity of a Sleeping Beauty (SB)‐based dual rAAV virion approach to achieve stable and efficient gene transfer to the newborn murine liver using transposable therapeutic cassettes encoding coagulation factor IX or ornithine transcarbamylase (OTC). Results: At equivalent doses, rAAV/SB100X transduced hepatocytes with high efficiency, achieving stable expression into adulthood. Compared with conventional AAV, the proportion of hepatocytes transduced, and factor IX and OTC activity levels, were both markedly increased. The proportion of hepatocytes stably transduced increased 4‐ to 8‐fold from <5%, and activity levels increased correspondingly, with markedly increased survival and stable urinary orotate levels in the OTC‐deficient Spfash mouse following elimination of residual endogenous murine OTC. Conclusions: The present study demonstrates the first in vivo utility of a hybrid rAAV/SB100X transposon system to achieve stable long‐term therapeutic gene expression following delivery to the highly proliferative newborn mouse liver. These results have relevance to the treatment of genetic metabolic liver diseases with neonatal onset. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. Sleeping BeautymRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency
- Author
-
Zakas, Philip M., Cunningham, Sharon C., Doherty, Ann, van Dijk, Eva B., Ibraheim, Raed, Yu, Stephanie, Mekonnen, Befikadu D., Lang, Brendan, English, Elizabeth J., Sun, Gang, Duncan, Miles C., Benczkowski, Matthew S., Altshuler, Robert C., Singh, Malvenderjit Jagjit, Kibbler, Emily S., Tonga, Gulen Y., Wang, Zi Jun, Wang, Z. Jane, Li, Guangde, An, Ding, Rottman, James B., Bhavsar, Yashvi, Purcell, Cormac, Jain, Rachit, Alberry, Ryan, Roquet, Nathaniel, Fu, Yanfang, Citorik, Robert J., Rubens, Jacob R., Holmes, Michael C., Cotta-Ramusino, Cecilia, Querbes, William, Alexander, Ian E., and Salomon, William E.
- Abstract
Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivodelivery approach based on an engineered transposase, Sleeping Beauty(SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spfashmouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types.
- Published
- 2024
- Full Text
- View/download PDF
4. An ingestible bacterial-electronic system to monitor gastrointestinal health
- Author
-
Mimee, Mark, Nadeau, Phillip, Hayward, Alison, Carim, Sean, Flanagan, Sarah, Jerger, Logan, Collins, Joy, McDonnell, Shane, Swartwout, Richard, Citorik, Robert J., Bulović, Vladimir, Langer, Robert, Traverso, Giovanni, Chandrakasan, Anantha P., and Lu, Timothy K.
- Published
- 2018
5. Bacteriophage Therapy Testing Against Shigella flexneri in a Novel Human Intestinal Organoid-Derived Infection Model
- Author
-
Llanos-Chea, Alejandro, Citorik, Robert J., Nickerson, Kourtney P., Ingano, Laura, Serena, Gloria, Senger, Stefania, Lu, Timothy K., Fasano, Alessio, and Faherty, Christina S.
- Published
- 2019
- Full Text
- View/download PDF
6. Efficient retroelement-mediated DNA writing in bacteria
- Author
-
Farzadfard, Fahim, Gharaei, Nava, Citorik, Robert J., Lu, Timothy K., Farzadfard, Fahim, Gharaei, Nava, Citorik, Robert J., and Lu, Timothy K.
- Abstract
The ability to efficiently and dynamically change information stored in genomes would enable powerful strategies for studying cell biology and controlling cellular phenotypes. Current recombineering-mediated DNA writing platforms in bacteria are limited to specific laboratory conditions, often suffer from suboptimal editing efficiencies, and are not suitable for in situ applications. To overcome these limitations, we engineered a retroelement-mediated DNA writing system that enables efficient and precise editing of bacterial genomes without the requirement for target-specific elements or selection. We demonstrate that this DNA writing platform enables a broad range of applications, including efficient, scarless, and cis-element-independent editing of targeted microbial genomes within complex communities, the high-throughput mapping of spatial information and cellular interactions into DNA memory, and the continuous evolution of cellular traits.
- Published
- 2022
7. LPLUNC1 Modulates Innate Immune Responses to Vibrio cholerae
- Author
-
Shin, Ok S., Uddin, Taher, Citorik, Robert, Wang, Jennifer P., Pelle, Patricia Della, Kradin, Richard L., Bingle, Colin D., Bingle, Lynne, Camilli, Andrew, Bhuiyan, Taufiqur R., Shirin, Tahmina, Ryan, Edward T., Calderwood, Stephen B., Finberg, Robert W., Qadri, Firdausi, LaRocque, Regina C., and Harris, Jason B.
- Published
- 2011
- Full Text
- View/download PDF
8. Synthetic Biology and Therapies for Infectious Diseases
- Author
-
Ando, Hiroki, primary, Citorik, Robert, additional, Cleto, Sara, additional, Lemire, Sebastien, additional, Mimee, Mark, additional, and Lu, Timothy, additional
- Published
- 2014
- Full Text
- View/download PDF
9. Efficient retroelement-mediated DNA writing in bacteria
- Author
-
Farzadfard, Fahim, primary, Gharaei, Nava, additional, Citorik, Robert J., additional, and Lu, Timothy K., additional
- Published
- 2021
- Full Text
- View/download PDF
10. Microbiome therapeutics — Advances and challenges
- Author
-
Massachusetts Institute of Technology. Microbiology Graduate Program, Massachusetts Institute of Technology. Synthetic Biology Center, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Mimee, Mark, Citorik, Robert J, Lu, Timothy K, Massachusetts Institute of Technology. Microbiology Graduate Program, Massachusetts Institute of Technology. Synthetic Biology Center, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Mimee, Mark, Citorik, Robert J, and Lu, Timothy K
- Abstract
© 2016 The microbial community that lives on and in the human body exerts a major impact on human health, from metabolism to immunity. In order to leverage the close associations between microbes and their host, development of therapeutics targeting the microbiota has surged in recent years. Here, we discuss current additive and subtractive strategies to manipulate the microbiota, focusing on bacteria engineered to produce therapeutic payloads, consortia of natural organisms and selective antimicrobials. Further, we present challenges faced by the community in the development of microbiome therapeutics, including designing microbial therapies that are adapted for specific geographies in the body, stable colonization with microbial therapies, discovery of clinically relevant biosensors, robustness of engineered synthetic gene circuits and addressing safety and biocontainment concerns. Moving forward, collaboration between basic and applied researchers and clinicians to address these challenges will poise the field to herald an age of next-generation, cellular therapies that draw on novel findings in basic research to inform directed augmentation of the human microbiota.
- Published
- 2021
11. A systems biology approach to modeling Vibrio cholerae gene expression under virulence-inducing conditions
- Author
-
Kanjilal, Sanjat, Citorik, Robert, LaRocque, Regina C., Ramoni, Marco F., and Calderwood, Stephen B.
- Subjects
Gene expression -- Models ,Systems biology -- Research ,Vibrio cholerae -- Genetic aspects ,Bacterial genetics -- Research ,Biological sciences - Abstract
Vibrio cholerae is a Gram-negative bacillus that is the causative agent of cholera. Pathogenesis in vivo occurs through a series of spatiotemporally controlled events under the control of a gene cascade termed the ToxR regulon. Major genes in the ToxR regulon include the master regulators toxRS and tcpPH, the downstream regulator toxT, and virulence factors, the ctxAB and tcpA operons. Our current understanding of the dynamics of virulence gene expression is limited to microarray analyses of expression at selected time points. To better understand this process, we utilized a systems biology approach to examine the temporal regulation of gene expression in El Tor V. cholerae grown under virulence-inducing conditions in vitro (AKI medium), using high-resolution time series genomic profiling. Results showed that overall gene expression in AKI medium mimics that of in vivo studies but with less clear temporal separation between upstream regulators and downstream targets. Expression of toxRS was unaffected by growth under virulence-inducing conditions, but expression of toxT was activated shortly after switching from stationary to aerating conditions. The tcpA operon was also activated early during mid-exponential-phase growth, while the ctxAB operon was turned on later, after the rise in toxT expression. Expression of ctxAB continued to rise despite an eventual decrease in toxT. Cluster analysis of gene expression highlighted 15 hypothetical genes and six genes related to environmental information processing that represent potential new members of the ToxR regulon. This study applies systems biology tools to analysis of gene expression of V. cholerae in vitro and provides an important comparator for future studies done in vivo. doi: 10.1128/JB.00182-10
- Published
- 2010
12. An ingestible bacterial-electronic system to monitor gastrointestinal health
- Author
-
Massachusetts Institute of Technology. Synthetic Biology Center, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Department of Chemical Engineering, Massachusetts Institute of Technology. Department of Biological Engineering, Koch Institute for Integrative Cancer Research at MIT, Mimee, Mark Kyle, Nadeau, Phillip, Hayward, Alison M, Carim, Sean, Flanagan, Sarah, Jerger, Logan Andrew, Collins, Joy E, McDonnell, Shane, Swartwout, Richard M, Citorik, Robert James, Bulovic, Vladimir, Langer, Robert S, Traverso, Carlo Giovanni, Chandrakasan, Anantha P, Lu, Timothy K, Massachusetts Institute of Technology. Synthetic Biology Center, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Department of Chemical Engineering, Massachusetts Institute of Technology. Department of Biological Engineering, Koch Institute for Integrative Cancer Research at MIT, Mimee, Mark Kyle, Nadeau, Phillip, Hayward, Alison M, Carim, Sean, Flanagan, Sarah, Jerger, Logan Andrew, Collins, Joy E, McDonnell, Shane, Swartwout, Richard M, Citorik, Robert James, Bulovic, Vladimir, Langer, Robert S, Traverso, Carlo Giovanni, Chandrakasan, Anantha P, and Lu, Timothy K
- Abstract
Biomolecular monitoring in the gastrointestinal tract could offer rapid, precise disease detection and management but is impeded by access to the remote and complex environment. Here, we present an ingestible micro-bio-electronic device (IMBED) for in situ biomolecular detection based on environmentally resilient biosensor bacteria and miniaturized luminescence readout electronics that wirelessly communicate with an external device. As a proof of concept, we engineer heme-sensitive probiotic biosensors and demonstrate accurate diagnosis of gastrointestinal bleeding in swine. Additionally, we integrate alternative biosensors to demonstrate modularity and extensibility of the detection platform. IMBEDs enable new opportunities for gastrointestinal biomarker discovery and could transform the management and diagnosis of gastrointestinal disease., Office of Naval Research (Grant N00014-13-1-0424), National Institutes of Health (Grant EB-000244)
- Published
- 2020
13. Efficient Retroelement-Mediated DNA Writing in Bacteria
- Author
-
Farzadfard, Fahim, primary, Gharaei, Nava, additional, Citorik, Robert J., additional, and Lu, Timothy K., additional
- Published
- 2020
- Full Text
- View/download PDF
14. Isolation of Lytic Bacteriophages against Multidrug-Resistant Enterococcus faecalis ST55 Isolated from Research Macaques
- Author
-
Lieberman, Mia T, Citorik, Robert J, Lu, Tim K, and Fox, James G
- Abstract
Objectives: Multidrug-resistant Enterococcus faecalis (MDR-Ef) is a major cause of human nosocomial infections1. We have identified MDR-Ef of sequence type (ST) 55 from multiple cephalic recording chambers of macaques used in neuroscience research. Whole genome sequencing identified genes encoding resistance to aminoglycosides, tetracycline, erythromycin, and macrolide antibiotics. Our aim was to isolate lytic bacteriophages with activity against MDR-Ef. Methodology: Sterile-filtered sewage samples were combined with double-strength phage broth and inoculated with one of seven ST55 isolates. Following overnight incubation, enriched samples were centrifuged and supernatants spot tested on ST55 isolates using a modified double-agar overlay method2. Individual plaques were purified and serially passaged three times prior to overnight polyethylene glycol precipitation, uranyl actetate staining and transmission electron microscopy (TEM). Results: Three isolates showed the strongest lytic activity against all seven ST55 isolates based on plaque size. Initial TEM imaging identified the three phages as likely Siphoviruses of two distinct morphologies, featuring long, flexible, apparently noncontractile tails. Capsid morphologies included icosahedral (~65nm diameter) and two sizes of prolate capsids (~99nm x 36nm and ~109nm x 44nm). Conclusions: Lytic phage therapy offers a valuable alternative to traditional antimicrobials against highly drug-resistant bacteria. Future work includes phage sequencing and characterization of anti-biofilm activity., Journal of Targeting Infectiuos Diseases, Vol 1, No 1 (2016)
- Published
- 2016
- Full Text
- View/download PDF
15. Synthetic Biogenesis of Bacterial Amyloid Nanomaterials with Tunable Inorganic–Organic Interfaces and Electrical Conductivity
- Author
-
Seker, Urartu Ozgur Safak, primary, Chen, Allen Y., additional, Citorik, Robert J., additional, and Lu, Timothy K., additional
- Published
- 2016
- Full Text
- View/download PDF
16. Synthesis and patterning of tunable multiscale materials with engineered cells
- Author
-
Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Microbiology Graduate Program, Massachusetts Institute of Technology. Research Laboratory of Electronics, Massachusetts Institute of Technology. Synthetic Biology Center, Chen, Allen Y., Deng, Zhengtao, Seker, Urartu O. S., Lu, Michelle Y., Citorik, Robert James, Zakeri, Bijan, Lu, Timothy K., Billings, Amanda N., Harvard University--MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Microbiology Graduate Program, Massachusetts Institute of Technology. Research Laboratory of Electronics, Massachusetts Institute of Technology. Synthetic Biology Center, Chen, Allen Y., Deng, Zhengtao, Seker, Urartu O. S., Lu, Michelle Y., Citorik, Robert James, Zakeri, Bijan, Lu, Timothy K., and Billings, Amanda N.
- Abstract
Many natural biological systems—such as biofilms, shells and skeletal tissues—are able to assemble multifunctional and environmentally responsive multiscale assemblies of living and non-living components. Here, by using inducible genetic circuits and cellular communication circuits to regulate Escherichia coli curli amyloid production, we show that E. coli cells can organize self-assembling amyloid fibrils across multiple length scales, producing amyloid-based materials that are either externally controllable or undergo autonomous patterning. We also interfaced curli fibrils with inorganic materials, such as gold nanoparticles (AuNPs) and quantum dots (QDs), and used these capabilities to create an environmentally responsive biofilm-based electrical switch, produce gold nanowires and nanorods, co-localize AuNPs with CdTe/CdS QDs to modulate QD fluorescence lifetimes, and nucleate the formation of fluorescent ZnS QDs. This work lays a foundation for synthesizing, patterning, and controlling functional composite materials with engineered cells., United States. Office of Naval Research, United States. Army Research Office, National Science Foundation (U.S.). Materials Research Science and Engineering Centers (Program) (Award DMR-0819762), Hertz Foundation, United States. Dept. of Defense, National Institutes of Health (U.S.). Medical Scientist Training Program (Grant T32GM007753), National Institute of Environmental Health Sciences (Training Grant in Toxicology 5 T32 ES7020-37), Presidential Early Career Award for Scientists and Engineers, National Institutes of Health (U.S.) (New Innovator Award 1DP2OD008435)
- Published
- 2016
17. Sequence-specific antimicrobials using efficiently delivered RNA-guided nucleases
- Author
-
Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Synthetic Biology Center, Citorik, Robert James, Mimee, Mark K., Lu, Timothy K., Mimee, Mark Kyle, Lu, Timothy K, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Synthetic Biology Center, Citorik, Robert James, Mimee, Mark K., Lu, Timothy K., Mimee, Mark Kyle, and Lu, Timothy K
- Abstract
Current antibiotics tend to be broad spectrum, leading to indiscriminate killing of commensal bacteria and accelerated evolution of drug resistance. Here, we use CRISPR-Cas technology to create antimicrobials whose spectrum of activity is chosen by design. RNA-guided nucleases (RGNs) targeting specific DNA sequences are delivered efficiently to microbial populations using bacteriophage or bacteria carrying plasmids transmissible by conjugation. The DNA targets of RGNs can be undesirable genes or polymorphisms, including antibiotic resistance and virulence determinants in carbapenem-resistant Enterobacteriaceae and enterohemorrhagic Escherichia coli. Delivery of RGNs significantly improves survival in a Galleria mellonella infection model. We also show that RGNs enable modulation of complex bacterial populations by selective knockdown of targeted strains based on genetic signatures. RGNs constitute a class of highly discriminatory, customizable antimicrobials that enact selective pressure at the DNA level to reduce the prevalence of undesired genes, minimize off-target effects and enable programmable remodeling of microbiota., National Institutes of Health (U.S.) (New Innovator Award 1DP2OD008435), National Centers for Systems Biology (U.S.) (Grant 1P50GM098792), United States. Defense Threat Reduction Agency (HDTRA1-14-1-0007), Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (W911NF13D0001), National Institute of General Medical Sciences (U.S.) (Interdepartmental Biotechnology Training Program 5T32 GM008334), Fonds de la recherche en sante du Quebec (Master's Training Award)
- Published
- 2016
18. Microbiome therapeutics — Advances and challenges
- Author
-
Mimee, Mark, primary, Citorik, Robert J., additional, and Lu, Timothy K., additional
- Published
- 2016
- Full Text
- View/download PDF
19. Bacteriophage Therapy Testing Against Shigella flexneriin a Novel Human Intestinal Organoid-Derived Infection Model
- Author
-
Llanos-Chea, Alejandro, Citorik, Robert J., Nickerson, Kourtney P., Ingano, Laura, Serena, Gloria, Senger, Stefania, Lu, Timothy K., Fasano, Alessio, and Faherty, Christina S.
- Abstract
Enteric bacterial pathogens cause diarrheal disease and mortality at significant rates throughout the world, particularly in children younger than 5 years. Our ability to combat bacterial pathogens has been hindered by antibiotic resistance, a lack of effective vaccines, and accurate models of infection. With the renewed interest in bacteriophage therapy, we sought to use a novel human intestinal model to investigate the efficacy of a newly isolated bacteriophage against Shigella flexneri. An S. flexneri2457T-specific bacteriophage was isolated and assessed through kill curve experiments and infection assays with colorectal adenocarcinoma HT-29 cells and a novel human intestinal organoid-derived epithelial monolayer model. In our treatment protocol, organoids were generated from intestinal crypt stem cells, expanded in culture, and seeded onto transwells to establish 2-dimensional monolayers that differentiate into intestinal cells. The isolated bacteriophage efficiently killed S. flexneri2457T, other S. flexneristrains, and a strain of 2457T harboring an antibiotic resistance cassette. Analyses with laboratory and commensal Escherichia colistrains demonstrated that the bacteriophage was specific to S. flexneri, as observed under co-culture conditions. Importantly, the bacteriophage prevented both S. flexneri2457T epithelial cell adherence and invasion in both infection models. Bacteriophages offer feasible alternatives to antibiotics for eliminating enteric pathogens, confirmed here by the bacteriophage-targeted killing of S. flexneri. Furthermore, application of the organoid model has provided important insight into Shigellapathogenesis and bacteriophage-dependent intervention strategies. The screening platform described herein provides proof-of-concept analysis for the development of novel bacteriophage therapies to target antibiotic-resistant pathogens.
- Published
- 2019
- Full Text
- View/download PDF
20. Bacteriophage-based synthetic biology for the study of infectious diseases
- Author
-
Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Synthetic Biology Center, Citorik, Robert James, Mimee, Mark K., Lu, Timothy K., Mimee, Mark Kyle, Lu, Timothy K, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Synthetic Biology Center, Citorik, Robert James, Mimee, Mark K., Lu, Timothy K., Mimee, Mark Kyle, and Lu, Timothy K
- Abstract
Since their discovery, bacteriophages have contributed enormously to our understanding of molecular biology as model systems. Furthermore, bacteriophages have provided many tools that have advanced the fields of genetic engineering and synthetic biology. Here, we discuss bacteriophage-based technologies and their application to the study of infectious diseases. New strategies for engineering genomes have the potential to accelerate the design of novel phages as therapies, diagnostics, and tools. Though almost a century has elapsed since their discovery, bacteriophages continue to have a major impact on modern biological sciences, especially with the growth of multidrug-resistant bacteria and interest in the microbiome., National Institutes of Health (U.S.) (New Innovator Award DP2 OD008435), National Institutes of Health (U.S.) (National Centers for Systems Biology Grant P50 GM098792), United States. Defense Threat Reduction Agency (022744-001), Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (W911NF13D0001), National Institute of General Medical Sciences (U.S.) (Interdepartmental Biotechnology Training Program 5T32 GM008334)
- Published
- 2014
21. Sequence-specific antimicrobials using efficiently delivered RNA-guided nucleases
- Author
-
Citorik, Robert J, primary, Mimee, Mark, additional, and Lu, Timothy K, additional
- Published
- 2014
- Full Text
- View/download PDF
22. Bacteriophage-based synthetic biology for the study of infectious diseases
- Author
-
Citorik, Robert J, primary, Mimee, Mark, additional, and Lu, Timothy K, additional
- Published
- 2014
- Full Text
- View/download PDF
23. Synthesis and patterning of tunable multiscale materials with engineered cells
- Author
-
Chen, Allen Y., primary, Deng, Zhengtao, additional, Billings, Amanda N., additional, Seker, Urartu O. S., additional, Lu, Michelle Y., additional, Citorik, Robert J., additional, Zakeri, Bijan, additional, and Lu, Timothy K., additional
- Published
- 2014
- Full Text
- View/download PDF
24. Synthesis and patterning of tunable multiscale materials with engineered cells
- Author
-
Chen, Allen Y, primary, Seker, Urartu O.S., additional, Lu, Michelle Y, additional, Citorik, Robert J, additional, and Lu, Timothy, additional
- Published
- 2014
- Full Text
- View/download PDF
25. Coming of phage.
- Author
-
Lu, Timothy and Citorik, Robert
- Subjects
- *
BACTERIOPHAGES , *ANTIBIOTICS , *PHARMACOKINETICS ,INFECTION treatment - Abstract
The article discusses the use of bacteriophages to treat infections instead of antibiotics. Topics include discovery of bacteriophages by English bacteriologist Frederick Twort; challenges of phage therapy and Phagoburn trial funded by the European Union and sponsored by Pherecydes Pharma. It also highlights the emergence of bacterial resistance to phages, bacterial strain coverage and pharmacokinetics.
- Published
- 2016
26. Sequence-specific antimicrobials using efficiently delivered RNA-guided nucleases
- Author
-
Timothy K. Lu, Mark Mimee, Robert J. Citorik, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology. Synthetic Biology Center, Citorik, Robert James, Mimee, Mark K., and Lu, Timothy K.
- Subjects
Population ,Biomedical Engineering ,Bioengineering ,Sequence (biology) ,Computational biology ,Applied Microbiology and Biotechnology ,Article ,Microbiology ,Bacteriophage ,Plasmid ,Ribonucleases ,Anti-Infective Agents ,Base sequence ,Bacteriophages ,education ,education.field_of_study ,biology ,Base Sequence ,RNA ,Gene targeting ,Drug Resistance, Microbial ,biology.organism_classification ,Antimicrobial ,Research Highlight ,humanities ,eye diseases ,Anti-Bacterial Agents ,Carbapenems ,Enterohemorrhagic Escherichia coli ,embryonic structures ,Gene Targeting ,Molecular Medicine ,CRISPR-Cas Systems ,human activities ,Biotechnology ,Plasmids ,RNA, Guide, Kinetoplastida - Abstract
Current antibiotics tend to be broad spectrum, leading to indiscriminate killing of commensal bacteria and accelerated evolution of drug resistance. Here, we use CRISPR-Cas technology to create antimicrobials whose spectrum of activity is chosen by design. RNA-guided nucleases (RGNs) targeting specific DNA sequences are delivered efficiently to microbial populations using bacteriophage or bacteria carrying plasmids transmissible by conjugation. The DNA targets of RGNs can be undesirable genes or polymorphisms, including antibiotic resistance and virulence determinants in carbapenem-resistant Enterobacteriaceae and enterohemorrhagic Escherichia coli. Delivery of RGNs significantly improves survival in a Galleria mellonella infection model. We also show that RGNs enable modulation of complex bacterial populations by selective knockdown of targeted strains based on genetic signatures. RGNs constitute a class of highly discriminatory, customizable antimicrobials that enact selective pressure at the DNA level to reduce the prevalence of undesired genes, minimize off-target effects and enable programmable remodeling of microbiota., National Institutes of Health (U.S.) (New Innovator Award 1DP2OD008435), National Centers for Systems Biology (U.S.) (Grant 1P50GM098792), United States. Defense Threat Reduction Agency (HDTRA1-14-1-0007), Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (W911NF13D0001), National Institute of General Medical Sciences (U.S.) (Interdepartmental Biotechnology Training Program 5T32 GM008334), Fonds de la recherche en sante du Quebec (Master's Training Award)
- Published
- 2014
27. Synthetic Biogenesis of Bacterial Amyloid Nanomaterials with Tunable Inorganic-Organic Interfaces and Electrical Conductivity.
- Author
-
Seker UO, Chen AY, Citorik RJ, and Lu TK
- Subjects
- Bacterial Proteins metabolism, Biofilms growth & development, Electric Conductivity, Escherichia coli Proteins metabolism, Gold metabolism, Metal Nanoparticles microbiology, Nanofibers, Nanotechnology methods, Nanowires microbiology, Particle Size, Amyloid metabolism, Escherichia coli metabolism, Nanostructures microbiology
- Abstract
Amyloids are highly ordered, hierarchal protein nanoassemblies. Functional amyloids in bacterial biofilms, such as Escherichia coli curli fibers, are formed by the polymerization of monomeric proteins secreted into the extracellular space. Curli is synthesized by living cells, is primarily composed of the major curlin subunit CsgA, and forms biological nanofibers with high aspect ratios. Here, we explore the application of curli fibers for nanotechnology by engineering curli to mediate tunable biological interfaces with inorganic materials and to controllably form gold nanoparticles and gold nanowires. Specifically, we used cell-synthesized curli fibers as templates for nucleating and growing gold nanoparticles and showed that nanoparticle size could be modulated as a function of curli fiber gold-binding affinity. Furthermore, we demonstrated that gold nanoparticles can be preseeded onto curli fibers and followed by gold enhancement to form nanowires. Using these two approaches, we created artificial cellular systems that integrate inorganic-organic materials to achieve tunable electrical conductivity. We envision that cell-synthesized amyloid nanofibers will be useful for interfacing abiotic and biotic systems to create living functional materials..
- Published
- 2017
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.