José A. Fernández Robledo, PhD


Senior Research Scientist
Molecular Biologist
Phone: +1 (207) 315-2567, ext. 315
Fax: +1 (207) 315-2329
jfernandez-robledo@bigelow.org

For media inquiries, please contact sprofaizer@bigelow.org



Education

Licentiate, Molecular Biology, U Santiago de Compostela, Spain

MS, Fish Pathology and Aquaculture, U Santiago de Compostela, Spain

PhD, Biochemistry and Molecular Biology, IIM-CSIC, U Santiago de Compostela, Spain


Research Interests

I am interested in a lot of things; I have experience in protozoan parasites of mollusks. My research aims to understanding the interaction between parasites and host. The system I have been using to study this interaction is Perkinsus marinus (‘Dermo’ disease) and Crassostrea virginica, the eastern oyster. Most of my scientific contribution has been on the side of the parasite where we have contributed to develop the genetic tools needed to make P. marinus a genetically traceable system. A spin off these projects is to use P. marinus to deliver antigens against pathogens of veterinarian and medical relevance (e.g., Malaria, SARS-CoV-2, VHSV). Prompted by limited intervention strategies to fight the spread of the Dermo, I am now exploring areas on the oyster side. With the genome of the oyster available and our recent achievement on transfection of the oyster hemocytes, I am interested in creating cell lines of oysters as a way to complete the circle for the study host-parasite interactions but also for exploring cellular aquaculture. I am also interested on using genetically engineering microalgae for ameliorating environmental problems and for producing food products. I am also looking for collaborations for bringing together AI to cell biology.

Funding

Logos: NSF, Gordon and Betty Moore Foundation, NOAA, NIH, Medicines for Malaria Venture

Molecular Parasitology and Genetic Engineering Group

On going

If you look for them, parasites are anywhere. We look for them in Maine, and indeed here they are. Unless someone comes with a new paradigm in immunology of invertebrates, as today, vaccination of oysters is not an option, since as invertebrates, oysters lack adaptive immunity; consequently, most intervention strategies have focused on management of the resource with variable success. Two of the major diseases that impact the oyster production in North America are “Dermo” disease caused by Perkinsus marinus and “MSX” (multinucleated sphere X) disease caused by Haplosporidium nelsoni. In Maine we have detected both of them with high prevalence and since they are here to stay, we are adapting to live with them. Often, parasites had a bad name and we often forget they have evolved to also play a role in the environment. Over the years, we have been working (I mean the post-docs and students) towards giving P. marinus a good name by developing the molecular tools necessary to accelerate their evolution and use them to solve pressing world health problems. For example, we have converted P. marinus in a platform, the first marine parasite, for expressing heterologous genes. We are focusing on vaccine candidate genes for Malaria, Cryptosporidiosis, and COVID-19. Within one week of receiving the plasmid containing the SARS-CoV-2 Spike gene, we have been able to express it in P. marinus. Our contribution to fight the modern “pest”. Thank the oysters the next time you are in a Raw Bar. Perkinsus marinus MOE

The bases for the development of the heterologous expression system is Perkinsus marinus MOE (PmMOE), a gene unique to the Perkinsus genus that lands on the surface of the trophozoite stage. Figure 01_MOE_1, 01_MOE_2, 01_MOE_3, 01_MOE_4

Heterologous expression in Perkinsus marinus

We are actively expressing genes of medical and veterinary relevance both to generate proteins for further characterization and as a vehicle for delivering antigens. Genes we are trying include the protozoan parasites Plasmodium falciparum, Toxoplasma gondii, Cryptosporidium hominis, and the virus Ebola, and SARS-CoV-2. Figure CSP, Figure COVID-19

On the make

As we are part of the environment, we are also interested in improving it without forgetting that we need to feed the planet. We already know there is a problem, talk to me about solutions. Our approach is to find ways to release pressure of our already exhausted resources at the same time that we create nutritious products and wealth for the people producing them. At the lab, we are exploring cellular aquaculture, a spin off from cellular agriculture. We ferment beer, wine, cheese, tofu, kimchi, Sriracha, why not grow seafood-derived products? We have genomes with thousands of genes, and nature and evolution are providing us with an increasing number of tools; we have to put them to work. This project is still clearing the field to take off. Figure 01_Cellular_aquaculture

Scope of Work

• Generate a repertoire of plasmids and genetic tools for continue engineering Perkinsus marinus

• As a genetic traceable protozoan explore the cell biology of Perkinsus marinus

• Apply the lessons learned making P. marinus a genetically traceable organism to other marine microeukaryotes

• Optimize P. marinus as a heterologous expression system and as new platform for delivering vaccines

• Identify stem cells in oyster and ways to create induced pluripotent stem cells (iPSC)

• Create cell lines from oysters

• Explore avenues for cellular aquaculture

• Keep educating students in responsible genetic engineering

• Using basic science to tackle environmental and health challenges

• Identifying ways to apply AI to cell biology

Publications

2013

  • Shridhar, S, Hassan, K, Sullivan, DJ, Vasta, GR, Fernández Robledo, JA. In press. Quantitative assessment of the proliferation of the protozoan parasite Perkinsus marinus using a bioluminescence assay for ATP content. Int J Parasitol: Drug Drug Resistance.

2012

  • Vasta, GR. Ahmed, H, Bianchet, MA, Fernández-Robledo, JA, Amzel, LM. 2012. Diversity in recognition of glycans by F-type lectins and galectins: Molecular, structural, and biophysical aspects. Annuals of the New York Academy of Sciences 1253, E14-26
  • Vasta, GR. Ahmed, H, Lazar, MN, Banerjee, A, Pasek, M, Shridhar, S, Guha, P, Fernández-Robledo, JA. 2012. Galectins as self/non-self recognition receptors in innate and adaptive immunity: An unresolved paradox. Frontiers in Immunology, 3: 199

2011

  • Fernández-Robledo, JA, Matsuzaki, M, Caler, Keeling, PJ, Roos, DS, Vasta, GR. 2011. The search for the missing link: a relic plastid in Perkinsus? International Journal for Parasitology. 41(12): 1217-1229
  • Lin, Z, Fernández-Robledo, JA, Cellier, MFM, Vasta, GR. 2011. The natural resistance-associated macrophage protein from the protozoan parasite Perkinsus marinus mediates iron uptake. Biochemistry,120(2): 257-268

2010

  • Fernández-Robledo JA, Vasta GR. 2010. Production of recombinant proteins from protozoan parasites. Trends in Parasitology, 26(5): 244-254
  • Joseph SJ*, Fernández-Robledo JA*, Gardner MJ, El-Sayed NM, Kuo C-H, Schott EJ, Wang, H, Kissinger JC, Vasta, G.R. 2010. The Alveolate Perkinsus marinus: Biological insights from EST gene discovery. BMC Genomics. 11: 228 * Contributed equally

2009

  • Lin, Z, Fernández-Robledo, JA, Cellier, MFM, Vasta, GR. 2009. Metals and membrane metal transporters in biological systems: The role(s) of Nramp in host-parasite interactions. Journal of the Argentinean Society of Chemistry. 97: 210-25
  • Alavi, MR, Fernández-Robledo, JA, Vasta, GR. 2009. In vitro Intracellular survival of Perkinsus marinus trophozoites upon phagocytosis by oyster (Crassostrea virginica and Crassostrea ariakensis) hemocytes. Journal of Parasitology. 95: 900-7

2008

  • Fernández-Robledo JA, Lin Z, Vasta GR. 2008. Transfection of the protozoan parasite Perkinsus marinus. Molecular and Biochemical Parasitology 157: 44-53
  • Fernández-Robledo JA, Schott EJ, Vasta GR. 2008. Perkinsus marinus superoxide dismutase 2 (PmSOD2) localizes to single-membrane subcellular compartments. Biochemical and Biophysical Research Communications 375: 215-9.
  • Pecher W, Alavi M, Schott E, Fernández-Robledo JA, Roth L, Berg S, et al. 2008. Assessment of the northern distribution range of selected Perkinsus species in eastern oysters (Crassostrea virginica) and hard clams (Mercenaria mercenaria) using PCR-based detection assays. Journal of Parasitology 94: 410–22
  • Schott EJ, Fernández-Robledo JA, Alavi MR, Vasta GR. 2008. Susceptibility of Crassostrea ariakensis (Fujita 1913) to Bonamia and Perkinsus spp. infections: potential for disease transmission between oyster species. Journal of Shellfish Research: 541–9

Prior to 2007

  • Pecher WT, Robledo JAF, Vasta GR. 2004. Identification of a second rRNA gene unit in the Perkinsus andrewsi genome. Journal of Eukaryotic Microbiology 51: 234-45
  • Robledo JAF, Courville P, Cellier MF, Vasta GR. 2004. Gene organization and expression of the divalent cation transporter Nramp in the protistan parasite Perkinsus marinus. Journal of Parasitology 90: 1004-14
  • Schott EJ, Robledo JAF, Wright AC, Silva AM, Vasta GR. 2003. Gene organization and homology modeling of two iron superoxide dismutases of the early branching protist Perkinsus marinus. Gene 309: 1-9
  • Quesenberry M, Saito K, Krupatkina D, Robledo JAF, Drgon T, Pecher W, et al. 2002. Bioassay for ichthyocidal activity of Pfiesteria piscicida: Characterization of a culture flask assay format. Journal of Applied Phycology 14: 241-54
  • Robledo JAF, Nunes PA, Cancela ML, Vasta GR. 2002. Development of an in vitro clonal culture and characterization of the rRNA gene cluster of Perkinsus atlanticus, a protistan parasite of the clam Tapes decussatus. Journal of Eukaryotic Microbiology 49: 414-22
  • Saito K, Drgon T, Robledo JAF, Krupatkina DN, Vasta GR. 2002. Characterization of the rRNA Locus of Pfiesteria piscicida and Development of Standard and Quantitative PCR-Based Detection Assays Targeted to the Nontranscribed Spacer. Applied and Environmental Microbiology 68: 5394-407
  • Coss CA, Robledo JAF, Ruiz GM, Vasta GR. 2001a. Description of Perkinsus andrewsi n. sp. Isolated from the Baltic clam (Macoma balthica) by Characterization of the Ribosomal RNA Locus, and Development of a Species-Specific PCR-Based Diagnostic Assay. Journal of Eukaryotic Microbiology 48: 52-61
  • Coss CA, Robledo JAF, Vasta GR. 2001b. Fine structure of clonally propagated in vitro life stages of a Perkinsus sp. isolated from the Baltic clam Macoma balthica. Journal of Eukaryotic Microbiology 48: 38-51
  • Robledo JAF, Coss CA, Vasta GR. 2000a. Characterization of the ribosomal RNA locus of Perkinsus atlanticus and development of a polymerase chain reaction-based diagnostic assay. Journal of Parasitology 86: 972-8.
  • Robledo JAF, Wright AC, Marsh AG, Vasta GR. 1999. Nucleotide sequence variability in the nontranscribed spacer of the rRNA locus in the oyster parasite Perkinsus marinus. Journal of Parasitology 85: 650-6
  • Robledo JAF, Gauthier JD, Coss CA, Wright AC, Vasta GR. 1998. Species-specificity and sensitivity of a PCR-based assay for Perkinsus marinus in the eastern oyster, Crassostrea virginica: a comparison with the fluid thioglycollate assay. Journal of Parasitology 84: 1237-44

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