Our research philosophy is based on a paradox notion that less is more. It refers to all aspects of the process (apart from funding) including team composition, the projects lineup, and, especially, the approaches.
Recognizing the inherent limitations of this philosophy, we expand our team’s expertise through collaborations at all levels, including UF-, country-, and worldwide. We try to stay nimble and follow (sometimes even lead) the most promising research trends and venues.
Recently, our major interests have aligned with the bioinformatics, machine learning (ML), and artificial intelligence (AI). Due to the current global circumstances, we were also mobilized to apply our bioinformatics and gene therapy expertise to the development of the anti-SARS-CoV-2 vaccine. Our main instrument is recombinant Adeno-associated virus (rAAV) vector, and the main focus of our research is identifying and developing more efficient capsids.
Our current research involves the application of artificial intelligence to improve AAV as a gene therapy vector. Specifically, we develop and use machine learning algorithms to interpret big data gathered through AAV library next-generation sequencing after following Directed Evolution. Potential areas of interest include AAV assembly, packaging, stability, tropism, and immune evasion.
In collaboration with Dr. Tran’s Laboratory , and using Directed Evolution, we identified novel AAV capsid specifically targeting human glioblastoma stem-like fast-, and slow proliferating cells.
In collaboration with Drs. D. Tran, W. G. Sawyer, and S. Eikenberry, and using AI developed to identify cell fate determinants, we’re working on enabling the reprogramming Mesenchymal Stem Cells (MSCs) into any desired cell lineage while by-passing traditional iPSCs stage.
A Historical and Current Perspective
Implementing a concept of significantly improving the expression of a transgene in a mammalian cell by utilizing synthetic cDNA sequence optimized for synonymous codons usage, as exemplified by designing a “humanized” sequence encoding A. victoria Green Fluorescent Protein (GFP).
The concept found its subsequent development in many other transgenes, including genes of mammalian origin, re-designed for the purpose of higher expression in a context of viral vectors produced for clinical trials in humans.
Implementing a concept of designing combinatorial synthetic capsid libraries for directed evolution of targeted adeno-associated virus (AAV) vectors.
The methodology was subsequently utilized by many laboratories around the world using different diversification strategies (error-prone PCR, family shuffling, and peptide display) to derive a variety of modified and tissue-targeted rAAV vectors.
Implementing novel methods of production and purification of recombinant AAV vectors irrespective of serotype.
In particular, a method of isotonic iodixanol density gradient is introduced allowing for one-step purification of different rAAV from crude lysate. Since its inception, this protocol is a method of choice for the whole gene therapy field utilizing rAAV vectors.
Designing a novel inducible system for a scale-up production of rAAV vectors in heterologous cells of insect origin infected with a single baculovirus vector.
This modular system is characterized by its utmost simplicity (consists of only two components), stability, and high yield of rAAV (exceeding previously known production systems by 10-fold).
Discovering an alternative satiation signaling pathway in the brain directly connecting oral cavity and hypothalamus.
This discovery describes novel approaches in therapy for obesity, a disease that so far has no pharmacological treatment solutions.