Juan Alfonso Redondo Marín

Summary

After obtaining a B. Sc. degree in Biology by the Complutense University of Madrid, with specialization in Biotechnology and Genetics, I carried out my postgraduate research in the Institute of Polymer Science and Technology (Spanish Research Council), focused in the evaluation of (meth)acrylic polymers as non-viral vectors for gene therapy. This work allowed me to obtain a Ph. D. in Chemistry by the Autónoma University of Madrid.

My predoctoral research was based on the hypothesis that chemical structure and charge distribution play an important role in the efficiency of cationic polymers as non-viral vectors. Cationic and neutral (meth)acrylic monomers were synthesized, characterized and copolymerized, obtaining linear copolymers that were evaluated in vitro as possible non-viral gene carriers . The results showed how the charge density and distribution plays an important role in the properties of the polyplexes (polymer/DNA complexes) and their transfection efficiency and cytocompatibility. Both parts of this interdisciplinar study, chemical synthesis and characterization, and the biological assays were carried out by me, so I got experience in basic molecular biology and cell culture techniques like in vitro transfection, viability assays and PCR, Western blot, and immunofluorescence, for example.

In addition, during my predoctoral stage, I also worked in the synthesis of conetworks using thermo and pH responsive polymers, in the biological evaluation of other biomaterials synthesized in my work group, and, finally, in the modification of natural polymers such as chitosan, and their evaluation as possible antitumoral biomaterials.

My postdoctoral research was developed in the Institute of Organic Chemistry and Biochemistry (Academy of Science of the Czech Republic) using in vitro selection to identify oligonucleotide sequences able to efficiently and specifically bind certain targets (so called aptamers) or efficiently catalyse biological reactions (so called (deoxy)ribozymes).

In particular, during my postdoctoral research, two in vitro selections were performed. The first selection was focused on finding an aptamer for the enzyme Glutamate carboxypeptidase III (GCPIII). This enzyme is closely related to Glutamate carboxypeptidase II (GCPII), which is involved in glutamatergic neurotoxicity and prostate cancer. The physiological role of GCPIII and its localization in humans/mice are unknown due to the lack of specific antibodies/ligands that can discriminate it from GCPII, due to the high homology between both enzymes.

The second in vitro selection looked for a ribozyme able to efficiently cleave single-stranded DNA. There are not natural (deoxy)ribozymes reported with the ability to efficiently cleave DNA under physiological conditions. Due to this reason, a ribozyme derived from the Group I Intron of Tetrahymena thermophila, able to cleave DNA under non-physiological conditions was picked as the ‘parent’ sequence for the selection. The sequence of this ribozyme was partially mutagenized obtaining an RNA pool that was subjected to in vitro selection under physiological K+ and Mg2+ concentrations.

As you can see, both my pre- and postdoctoral research are focused on gene therapy, either by creating the ‘vehicle’ to deliver the DNA, or by looking for the specific RNA or DNA sequence that could be the ‘active agent’. All of my  work was carried out in an interdisciplinary environment and I enjoy interacting with colleagues from different fields. I endeavor to learn and share from/with other researchers to apply scientific research to the ‘real world’.

Positions

Education

Skills

Monomer synthesis and polymerization techniques including controlled living polymerization.

Polymer characterization including GPC analysis and NMR.

Development of novel smart polymer systems for controlled delivery purposes.

Cell culture and biomaterial biocompatibility assessment using a variety of cell lines and quantification assays (Alamar Blue, MTS, calcein).

Genetic Engineering, Western Blot, protein purification.

Design, assembly and purifcation of oligonucleotides.

PCR, RT-PCR, Mutagenic PCR, T7 DNA transcription...

Cloning and sequencing of DNA oligonucleotides

Detection and purificatoin of nucleic acid/protein complexes or non-canonical DNA conformations using magnetic beads or native PAGE electrophoresis (EMSA).

Handling and management of radioactive (32P) material. Radiolabeling of oligonucletides.

CV

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