A5083707286- Genomics and Phylogenetic Studies
- Virus-based gene therapy research
- Microbial Community Ecology and Physiology
- Viral Infectious Diseases and Gene Expression in Insects
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Methane Hydrates and Related Phenomena
- Retinal Development and Disorders
- Polar Research and Ecology
- Bacterial Identification and Susceptibility Testing
- Viral Infections and Immunology Research
- Animal Genetics and Reproduction
- Retinal Diseases and Treatments
- RNA and protein synthesis mechanisms
- Planetary Science and Exploration
- Retinal and Optic Conditions
- CAR-T cell therapy research
- Biosensors and Analytical Detection
- Spaceflight effects on biology
- Space Exploration and Technology
- Paleontology and Stratigraphy of Fossils
- Enzyme Production and Characterization
- Food Allergy and Anaphylaxis Research
- SARS-CoV-2 and COVID-19 Research
- Mine drainage and remediation techniques
University of Florida
2014-2024
Florida College
2017-2022
University of Alabama in Huntsville
1999-2009
HudsonAlpha Institute for Biotechnology
2008
Lawrence Berkeley National Laboratory
2008
Alabama Agricultural and Mechanical University
2002
The major drawback of the Baculovirus/Sf9 system for recombinant adeno-associated viral (rAAV) manufacturing is that most Bac-derived rAAV vector serotypes, with few exceptions, demonstrate altered capsid compositions and lower biological potencies. Here, we describe a new insect cell-based production platform utilizing attenuated Kozak sequence leaky ribosome scanning to achieve serotype-specific modulation AAV proteins stoichiometry. By way example, rAAV5 rAAV9 were produced...
Methodologies to improve existing adeno-associated virus (AAV) vectors for gene therapy include either rational approaches or directed evolution derive capsid variants characterized by superior transduction efficiencies in targeted tissues. Here, we integrated both one unified design strategy of "virtual family shuffling" a combinatorial library whereby only variable regions on the surface are modified. Individual sublibraries were first assembled order preselect compatible amino acid...
A novel alkaliphilic, sulfate-reducing bacterium, strain MLF1(T), was isolated from sediments of soda Mono Lake, California. Gram-negative vibrio-shaped cells were observed, which 0.6-0.7x1.2-2.7 micro m in size, motile by a single polar flagellum and occurred singly, pairs or as short spirilla. Growth observed at 15-48 degrees C (optimum, 37 C), >1-7 % NaCl, w/v 3 %) pH 8.0-10.0 9.5). The isolate is strictly requires high concentration carbonate the growth medium obligately anaerobic...
A novel, obligately anaerobic, mesophilic, haloalkaliphilic spirochaete, strain ASpG1T, was isolated from sediments of the alkaline, hypersaline Mono Lake in California, USA. Cells Gram-negative were motile and spirochaete-shaped with sizes 0·2–0·22×8–18 μm. Growth observed between 10 44 °C (optimum 37 °C), 2–12 % (w/v) NaCl 3 NaCl) pH 8 10·5 9·5). The novel strictly alkaliphilic, required high concentrations carbonates medium capable utilizing d-glucose, fructose, maltose, sucrose, starch...
A novel, obligately anaerobic, psychrotolerant bacterium, designated strain PPP2T, was isolated from guano of the Magellanic penguin (Spheniscus magellanicus) in Chilean Patagonia. Cells were Gram-stain-positive, spore-forming, straight rods (0.7–0.8×3.0–5.0 μm) that motile by means peritrichous flagella. Growth observed at pH 6.7−9.7 (optimum 8.3) and 2–37 °C 29 °C). between 0 4 % (w/v) NaCl with optimum growth 0.5 (w/v). Strain PPP2T a catalase-negative chemo-organoheterotroph capable...
A novel, psychrotolerant, facultative anaerobe, strain FTR1 T , was isolated from Pleistocene ice the permafrost tunnel in Fox, Alaska. Gram-positive, motile, rod-shaped cells were observed with sizes 0·6–0·7×0·9–1·5 μm. Growth occurred within pH range 6·5–9·5 optimum growth at 7·3–7·5. The temperature for of novel isolate 0–28 °C and 24 °C. does not require NaCl; between 0 5 % NaCl 0·5 (w/v). a catalase-negative chemoorganoheterotroph that used as substrates sugars some products...
A novel, extremely psychrotolerant, facultative anaerobe, strain PmagG1 T , was isolated from guano of Magellanic penguins ( Spheniscus magellanicus ) collected in Chilean Patagonia. Gram-variable, motile cocci with a diameter 1.3–2.0 μm were observed singularly or pairs, short chains and irregular conglomerates. Growth occurred within the pH range 6.0–10.0, optimum growth at 8.5. The temperature for novel isolate −5 to 35 °C, 28–30 °C. Strain did not require NaCl, as presence 0–6.5 % NaCl...
A hyperthermophilic, sulfur-reducing, organo-heterotrophic archaeon, strain OGL-20P(T), was isolated from 'black smoker' chimney material the Rainbow hydrothermal vent site on Mid-Atlantic Ridge (36.2 degrees N, 33.9 W). The cells of OGL-20P(T) have an irregular coccoid shape and are motile with a single flagellum. Growth observed within pH range 5.0-8.5 (optimum 7.0), NaCl concentration 1-5 % (w/v) 3 %) temperature 55-94 C 83-85 C). novel isolate is strictly anaerobic obligately dependent...
Biodistribution analysis is a key step in the evaluation of adeno-associated virus (AAV) capsid variants, whether natural isolates or produced by rational design directed evolution. Indeed, when screening candidate vectors, accurate knowledge about which tissues are infected and how efficiently essential. We describe design, validation, application new vector, pTR-UF50-BC, encoding bioluminescent protein, fluorescent protein DNA barcode, can be used to visualize localization transduction at...
Limitations to successful gene therapy with adeno-associated virus (AAV) can comprise pre-existing neutralizing antibodies the vector capsid that block cellular entry, or inefficient transduction of target cells lead sub-optimal expression therapeutic transgene. Recombinant serotype 3 AAV (AAV3) is an emerging candidate for liver-directed therapy. In this study, we integrated rational design by using a combinatorial library derived from AAV3B capsids directed evolution in vitro selection...
Alpha-1 antitrypsin deficiency (AATD) is characterized by both chronic lung disease due to loss of wild-type AAT (M-AAT) antiprotease function and liver toxicity from delayed secretion, polymerization, aggregation misfolded mutant (Z-AAT). The ideal gene therapy for AATD should therefore comprise endogenous Z-AAT suppression M-AAT overexpression. We designed a dual-function rAAV3B (df-rAAV3B) construct, which was effective at transducing hepatocytes, resulting in considerable decrease levels...
A novel obligately anaerobic, mesophilic, alkaliphilic spirochaete, strain ASpC2T, was isolated from an anaerobic sediment of alkaline, hypersaline Owens Lake in California, USA. The Gram-negative cells are motile, helical shape and 0.23×8.0–18.0 μm. Growth occurs within the following ranges: 13–41 °C, with optimal growth at 35 °C; 1–3 % (w/v) NaCl, 2 NaCl; pH 7.8–10.5, 10.0. isolate is strictly requires high concentrations carbonate ions medium. It utilizes some sugars, organic acids, amino...
A novel obligately anaerobic, mesophilic, alkaliphilic spirochaete, strain ASpC2T, was isolated from an anaerobic sediment of alkaline, hypersaline Owens Lake in California, USA. The Gram-negative cells are motile, helical shape and 0.23×8.0–18.0 μm. Growth occurs within the following ranges: 13–41 °C, with optimal growth at 35 °C; 1–3 % (w/v) NaCl, 2 NaCl; pH 7.8–10.5, 10.0. isolate is strictly requires high concentrations carbonate ions medium. It utilizes some sugars, organic acids, amino...
Abstract Background Gene synthesis technologies are an important tool for structural biology projects, allowing increased protein expression through codon optimization and facilitating sequence alterations. Existing methods, however, can be complex not always reproducible, prompting researchers to use commercial suppliers rather than synthesize genes themselves. Results A PCR-based gene method, referred as SeqTBIO, is described efficiently assemble the coding regions of two novel...
Adeno-associated virus (AAV) gene therapy vectors have shown the best outcomes in human clinical studies for treatment of genetic diseases such as hemophilia. However, these pivotal investigations also identified several challenges. For example, high vector doses are often used hepatic transfer, and cytotoxic T lymphocyte responses against viral capsid may occur. Therefore, achieving at reduced other strategies to reduce antigen presentation desirable.We tested engineered AAV capsids factor...
Adeno-associated virus (AAV) continues to be the gold standard vector for therapeutic gene delivery and has proven especially useful treating ocular disease. Intravitreal injection (IVtI) is a promising route because it increases accessibility of therapies larger patient populations. However, data from clinical non-human primate (NHP) studies utilizing currently available capsids indicate that anatomical barriers AAV pre-existing neutralizing antibodies can restrict expression levels are...
Multiple independent adeno-associated virus (AAV) gene therapy clinical trials for hemophilia B, utilizing different AAV serotypes, have reported a vector dose-dependent loss of circulating factor IX (FIX) protein associated with capsid-specific CD8+ T cell (Cap-CD8) elimination transduced hepatocytes. Hemophilia B patients who develop transient transaminitis and FIX may be stabilized the immune-suppressive (IS) drug prednisolone, but do not all recover lost expression, whereas some fail to...
Capsid engineering of adeno-associated virus (AAV) can surmount current limitations to gene therapy such as broad tissue tropism, low transduction efficiency, or pre-existing neutralizing antibodies (NAb) that restrict patient eligibility. We previously generated an AAV3B combinatorial capsid library by integrating rational design and directed evolution with the aim improving hepatotropism. A potential isolate, AAV3B-DE5, gained a selective proliferative advantage over five rounds iterative...
Peanut is one of the most allergenic foods and reports accidental ingestion peanuts in unsuspected food are increasing. No information available on allergen content peanut germplasm grown commercially used confectionery industry. The objectives this study were: (1) to determine contents 34 introductions (PI); (2) identify naturally occurring allergen-free and/or low or hypoallergenic germplasm. A basic ELISA protocol was utilized detect presence antigens lines using a pool human sera from...
Purpose: Retinal pericytes play a vital role in maintaining retinal homeostasis, and their dysfunction underlies pathogenesis such vascular eye diseases as diabetic retinopathy wet age-related macular degeneration. Consequently, are attractive therapeutic targets for gene therapy, but effectively targeting with recombinant adeno-associated virus (rAAV) vectors remains challenge. Methods: We introduced genetic modifications into the surface-exposed variable regions of rAAV2/2 capsid to...
The bacterial diversity of microbial extremophiles from the meromictic, hypersaline Mono Lake and a small evaporite pool in Owens California was studied. In spite these regions had differing mineral background different concentrations NaCl water they contain same halo- alkaliphiles anaerobic community. Three new species bacteria were detected this community: primary anaerobe, dissipotrophic saccharolytic spirochete <i>Spirochaeta americana </i>strain AspG1<sup>T</sup>, anaerobe which is...
Retinal diseases are an important focus of in vivo gene therapy, for which adeno-associated virus is a preferred vector. Intravitreal injection highly desirable delivery route because it much simpler, safer and inexpensive than the main alternative subretinal injection. However, existing capsids have low efficiency, require high doses susceptible to neutralization by antibodies that present vitreous. Using AAV5 as starting material, we applied combination directed evolution rational design...