Molecular Biology
The Molecular Biology Unit (UBM) offers standardized nucleic acid sequencing services and fragment analysis by capillary electrophoresis. These services are aimed to provide technical support and scientific advice to researchers of the institute and other public or private research organizations, companies and centers across the region.
It also offers advice for the experimental design of genomic and transcriptomic studies through DNA microarrays or massive sequencing. It has a microarray scanner (Agilent) and a high capacity sequencer (Illumina) that allows the realization of a wide spectrum of experiments.
Members
Lisvane Paes, PhD
Senior Research Assistant
lpaes@pasteur.edu.uy
María Laura Chiribao, PhD
Honorary research associate
Postdoc
Faculty of Medicine, Udelar
chiribao@pasteur.edu.uy
Florencia Ruppel, DVM
Masters student
fruppel@pasteur.edu.uy
Research lines
Study of the intracellular host-pathogen interaction with a systemic approach
At the lab we have studied virulence factors of intracellular pathogens, as well as the host genes and routes necessary for the establishment of the infection, and the interface between them. We set out as main objectives the design of new strategies for the treatment or prevention of various infectious pathologies, such as Chagas disease, African trypanosomiasis, leishmaniasis, and tuberculosis
Determination of virulence factors necessary for T.cruzi infection
Regarding the establishment and persistence of T. cruzi infection, we have described a set of proteins related to the redox metabolism of trypanosomatids (triparedoxin cytosolic and mitochondrial peroxidase, triparredoxins, glutarredoxins, pteridin reductase), and we have characterized their structure and function. We are developing strategies for its inhibition, as well as its application in the development of therapeutic and preventive strategies.
Characterization of changes in gene expression produced by intracellular parasites in human cells
We have shown that there is a cellular reprogramming led by T. cruzi that allows the establishment and persistence of the infection in human cells. One of the main objectives we are currently pursuing is to identify host proteins, which, when inhibited, prevent T. cruzi infection. We are also extending this strategy for genomic and molecular biology studies in leishmaniasis, African trypanosomiasis, neosporosis and tuberculosis.
Services
DNA SEQUENCING (SANGER METHOD)
The UBM has the only facility in Uruguay for DNA sequencing service that performs sequencing using the Sanger Method and fragment analysis. We are working with a Genetic Analyzer 3130 (Applied BioSystems), and recently we acquired a 3500 Genetic Analyzer (Applied BioSystems), which will improve performance. The unit receives samples from private and public institutions throughout the country.
Bellow there will be found the cost of the services provided by the unit:
| SERVICE | COST (US$)* | COUPON BOOK x100 (US$)* | PRIVATE |
| Sequence | 7 | 600 | 20 |
| Fragments | 2,5 | 200 | 5 |
| Purification | 2,5 | 200 | 5 |
* Prices for academic sector (national and international)
The coupons expires 12 months after purchase.
- For the purchase of services, please write to: tecnologia@pasteur.edu.uy
- For consultation techniques or other services, please write to: ubm@pasteur.edu.uy
- To submit samples it is necessary to complete the following forms: Sequence Request Form or Fragment Analysis Form.
MICROARRAYS
The Agilent platform is still being used for transcriptomic studies of complex organisms, mainly human and bovine samples. There is also sporadic cooperation with private clinics that use Comparative Genomic Hybridization in diagnostics (for example, genetic diagnoses prior to implantation).
DEEP SEQUENCING
The Illumina device has led to the beginning of the Genomics Program, and is now widely used in different applications, mainly genomics and transcriptomics of pathogens (bacteria and unicellular protozoa). We also provide practical training on how to use the equipment and analyze the data, in order to stimulate the autonomy of other laboratories in the application of next-generation sequencing in their research.
Equipment
- DNA sequencer / analyzer
- Real-time PCR
- Microarray reader
- Microarray hybridizer
- BioAnalyzer
- General molecular biology instrumentation
- MiSeq Illumina
Courses
- “Functional Genomics and its applications in biomedicine: Host-Pathogen interaction”. RIIP/UNU-Biolac Course. November 2014.
Main publications
vacio
2018
- Díaz-Viraqué F, Chiribao ML, Trochine A, Gonzalez F, Castillo C,Liempi A, Kemmerling U, Maya JM, Robello C. (2018). Old Yellow Enzyme from Trypanosoma cruzi exhibits in vivo prostaglandin F2α synthase activity and has a key role in parasite infection and drug susceptibility. Frontiers in Immunology, 9, 456. doi:10.3389/fimmu.2018.00456
- Libisch M.G, Faral-Tello P, J.Garg N, Radi R, Piacenza L, Robello C.(2018) Early Trypanosoma cruzi Infection Triggers mTORC1-Mediated Respiration Increase and Mitochondrial Biogenesis in Human Primary Cardiomyocytes. Frontiers in Microbiology.9,1889.doi: 10.3389/fmicb.2018.01889
- Berná L, Rodriguez M, Chiribao ML, Parodi-Talice A, Pita S, Rijo G, Alvarez-Valin F, Robello C. Expanding an expanded genome: long-read sequencing of Trypanosoma cruzi. Microb Genom. 2018 Apr 30. doi: 10.1099/mgen.0.000177.
- Cavalieri D, Di Paola M, Rizzetto L, Tocci N, De Filippo C, Lionetti P, Ardizzoni A, Colombari B, Paulone S, Gut IG, Berná L, Gut M, Blanc J, Kapushesky M, Pericolini E, Blasi E, Peppoloni S. Genomic and Phenotypic Variation in Morphogenetic Networks of Two Candida albicans Isolates Subtends Their Different Pathogenic Potential. Front Immunol. 2018 Jan 19;8:1997. doi: 10.3389/fimmu.2017.01997
- Lasserre M, Fresia P, Greif G, Iraola G, Castro-Ramos M, Juambeltz A, Nuñez Á, Naya H, Robello C, Berná L. Whole genome sequencing of the monomorphic pathogen Mycobacterium bovis reveals local differentiation of cattle clinical isolates. BMC Genomics. 2018 Jan 2;19(1):2. doi: 10.1186/s12864-017-4249-6.
2017
- Greif G, Faral-Tello P, Scardoelli Vianna C, Hernandez A, Basmadjian Y, Robello C. The first case report of trypanosomiasis caused by Trypanosoma evansi in Uruguay. Veterinary Parasitology: Regional Studies and Reports. 2017 Nov; 11 (2017) 19–21. Doi: https://doi.org/10.1016/j.vprsr.2017.11.002
- Varela J, Birriel E, Nargoli J, Faral-Tello P, Robello C, Coqueiro A, Hae Choi Y, Cerecetto H, González M. (2017) Identification of New Anti-Trypanosoma Cruzi Agents in Some Uruguayan Plants by NMR-Based Metabolomic Profiling. Arch Nat Med Chem 2017: ANMC-105.
- Faral-Tello P, Satragno D, Canneva B, Verger L, Lozano A, Vitale E, Greif G, Soto C, Robello C, Basmadjián Y. Autochthonous Outbreak and Expansion of canine Visceral Leishmaniasis, Uruguay. Emerg Infect Dis. 2017 Mar; 23(3):536-538. doi:10.3201/eid2303.160377.
- Festari M.F., Trajtenberg F., Berois N., Pantano S., Revoredo L., Kong Y., Solari-Saquieres P., Narimatsu Y., Freire T., Bay S., Robello C., Bénard J., Gerken T.A., Clausen H., Osinaga E. 2017. Revisiting the human polypeptide GalNAc-T1 and T13 paralogs. Glycobiology. 27(1):140-153
- Berná L., Chiribao ML,Greif G, Rodriguez M, Alvarez-Valin F, Robello C. Transcriptomic analysis reveals metabolic switches and surface remodeling as key processes for stage transition in Trypanosoma cruzi. PeerJ. 2017 Mar 8;5:e3017. doi: 10.7717/peerj.3017. eCollection 2017.
- de Oliveira TC, Rodrigues PT, Menezes MJ, Gonçalves-Lopes RM, Bastos MS, Lima NF, Barbosa S, Gerber AL, Loss de Morais G, Berná L, Phelan J, Robello C, de Vasconcelos ATR, Alves JMP, Ferreira MU. Genome-wide diversity and differentiation in New World populations of the human malaria parasite Plasmodium vivax. PLoS Negl Trop Dis. 2017 Jul 31;11(7):e0005824. doi: 10.1371/journal.pntd.0005824. eCollection 2017 Jul.
2016
- Pizzo C, Faral-Tello P, Yaluff G, Serna E,Torres S, Vera N, Saiz C, Robello C, Mahler G. New approach towards the synthesis of selenosemicarbazones, usefulcompounds for Chagas’ disease. Eur J Med Chem. 2016 Feb 15;109:107-13.
- Ubillos L, Freire T, Berriel E, Chiribao ML, Chiale C, Festari MF, Medeiros A,Mazal D, Rondán M, Bollati-Fogolín M, Rabinovich GA, Robello C, Osinaga E. Trypanosoma cruzi extracts elicit protective immune response against chemicallyinduced colon and mammary cancers. Int J Cancer. 2016 Apr 1;138(7):1719-31
2015
- Coitinho C, Greif G, van Ingen J, Laserra P, Robello C, Rivas C. First case of Mycobacterium heckeshornense cavitary lung disease in the Latin America and Caribbean region. New Microbes New Infect. 2015 Dec 18;9:63-5.
- Dusfour I, Zorrilla P, Guidez A, Issaly J, Girod R, Guillaumot L, Robello C, Strode C. Deltamethrin Resistance Mechanisms in Aedes aegypti Populations fromThree French Overseas Territories Worldwide. PLoS Negl Trop Dis. 2015 Nov20;9(11).
- Lasserre M, Berná L, Greif G, Díaz-Viraqué F, Iraola G, Naya H, Castro-Ramos M, Juambeltz A, Robello C. Whole-Genome Sequences of Mycobacterium bovis StrainMbURU-001, Isolated from Fresh Bovine Infected Samples. Genome Announc. 2015 Nov 5;3(6).
- Greif G, Rodriguez M, Reyna-Bello A, Robello C, Alvarez-Valin F. Kinetoplast adaptations in American strains from Trypanosoma vivax. Mutat Res. 2015 Mar;773:69-82.
- Fernandez-Calero T, Garcia-Silva R, Pena A, Robello C, Persson H, Rovira C, Naya H, Cayota A. Profiling of small RNA cargo of extracellular vesicles shed by Trypanosoma cruzi reveals a specific extracellular signature. Mol Biochem Parasitol. 2015 Jan-Feb;199(1-2):19-28.
- Arias DG, Piñeyro MD, Iglesias AA, Guerrero SA, Robello C. Molecular characterization and interactome analysis of Trypanosoma cruzi tryparedoxin II. JProteomics. 2015 Apr 29;120:95-104.
- Trochine A, Creek DJ, Faral-Tello P, Barrett MP, Robello C. Bestatin induces specific changes in Trypanosoma cruzi dipeptide pool. Antimicrob Agents Chemother. 2015 May;59(5):2921-5.
- Querido JF, Echeverría MG, Marti GA, Costa RM, Susevich ML, Rabinovich JE, Copa A, Montaño NA, Garcia L, Cordova M, Torrico F, Sánchez-Eugenia R, Sánchez-Magraner L, Muñiz-Trabudua X, López-Marijuan I, Rozas-Dennis GS, Diosque P, de Castro AM, Robello C, Rodríguez JS, Altcheh J, Salazar-Schettino PM, Bucio MI, Espinoza B, Guérin DM, Silva MS. Seroprevalence of Triatoma virus (Dicistroviridae: Cripaviridae) antibodies in Chagas disease patients. Parasit Vectors. 2015 Jan 17;8:29.
- Palacios F, Abreu C, Prieto D, Morande P, Ruiz S, Fernández-Calero T, Naya H, Libisch G, Robello C, Landoni AI, Gabus R, Dighiero G, Oppezzo P. Activation ofthe PI3K/AKT pathway by microRNA-22 results in CLL B-cell proliferation. Leukemia. 2015 Jan;29(1):115-25.
2014
- Márquez VE, Arias DG, Chiribao ML, Faral-Tello P, Robello C, Iglesias AA, Guerrero SA. Redox metabolism in Trypanosoma cruzi. Biochemical characterization of dithiol glutaredoxin dependent cellular pathways. Biochimie. 2014Nov;106:56-67.
- Berná L, Iraola G, Greif G, Coitinho C, Rivas CM, Naya H, Robello C. Whole-Genome Sequencing of an Isoniazid-Resistant Clinical Isolate ofMycobacterium tuberculosis Strain MtURU-002 from Uruguay. Genome Announc. 2014Jul 17;2(4). pii: e00655-14.
- Giorello FM, Berná L, Greif G, Camesasca L, Salzman V, Medina K, Robello C, Gaggero C, Aguilar PS, Carrau F. Genome Sequence of the Native Apiculate Wine Yeast Hanseniaspora vineae T02/19AF. Genome Announc. 2014 May 29;2(3). pii:e00530-14.
- Trochine A, Creek DJ, Faral-Tello P, Barrett MP, Robello C. Benznidazole biotransformation and multiple targets in Trypanosoma cruzi revealed bymetabolomics. PLoS Negl Trop Dis. 2014 May 22;8(5):e2844.
- Chiribao ML, Libisch G, Parodi-Talice A, Robello C. Early Trypanosoma cruzi infection reprograms human epithelial cells. Biomed Res Int. 2014;2014:439501.
- Trochine A, Alvarez G, Corre S, Faral-Tello P, Durán R, Batthyany CI, Cerecetto H, González M, Robello C. Trypanosoma cruzi chemical proteomics using immobilized benznidazole. Exp Parasitol. 2014 May;140:33-8.
- Martinez A, Peluffo G, Petruk AA, Hugo M, Piñeyro D, Demicheli V, Moreno DM, Lima A, Batthyány C, Durán R, Robello C, Martí MA, Larrieux N, Buschiazzo A, Trujillo M, Radi R, Piacenza L. Structural and molecular basis of theperoxynitrite-mediated nitration and inactivation of Trypanosoma cruziiron-superoxide dismutases (Fe-SODs) A and B: disparate susceptibilities due tothe repair of Tyr35 radical by Cys83 in Fe-SODB through intramolecular electrontransfer. J Biol Chem. 2014 May 2;289(18):12760-78.
- Greif G, Iraola G, Berná L, Coitinho C, Rivas CM, Naya H, Robello C. Complete Genome Sequence of Mycobacterium tuberculosis Strain MtURU-001, Isolated from a Rapidly Progressing Outbreak in Uruguay. Genome Announc. 2014 Jan 23;2(1). pii:e01220-13.
- Faral-Tello P, Liang M, Mahler G, Wipf P, Robello C. Imidazolium compounds are active against all stages of Trypanosoma cruzi. Int J Antimicrob Agents. 2014 Mar;43(3):262-8.
- Garcia-Silva MR, das Neves RF, Cabrera-Cabrera F, Sanguinetti J, Medeiros LC, Robello C, Naya H, Fernandez-Calero T, Souto-Padron T, de Souza W, Cayota A. Extracellular vesicles shed by Trypanosoma cruzi are linked to small RNApathways, life cycle regulation, and susceptibility to infection of mammalian cells. Parasitol Res. 2014 Jan;113(1):285-304.
- Coitinho C, Greif G, Robello C, Laserra P, Willery E, Supply P. Rapidly progressing tuberculosis outbreak in a very low risk group. Eur Respir J. 2014 Mar;43(3):903-6.
- Libisch MG, Casás M, Chiribao M, Moreno P, Cayota A, Osinaga E, Oppezzo P, Robello C. GALNT11 as a new molecular marker in chronic lymphocytic leukemia. Gene. 2014 Jan 1;533(1):270-9.
2013
- Arias DG, Marquez VE, Chiribao ML, Gadelha FR, Robello C, Iglesias AA, Guerrero SA. Redox metabolism in Trypanosoma cruzi: functional characterization of tryparedoxins revisited. Free Radic Biol Med. 2013 Oct;63:65-77.
- Greif G, Ponce de Leon M, Lamolle G, Rodriguez M, Piñeyro D, Tavares-Marques LM, Reyna-Bello A, Robello C, Alvarez-Valin F. Transcriptome analysis of the bloodstream stage from the parasite Trypanosoma vivax. BMC Genomics. 2013 Mar5;14:149.
- Gadelha FR, Gonçalves CC, Mattos EC, Alves MJ, Piñeyro MD, Robello C, Peloso EF. Release of the cytosolic tryparedoxin peroxidase into the incubation medium and a different profile of cytosolic and mitochondrial peroxiredoxin expression in H2O2-treated Trypanosoma cruzi tissue culture-derived trypomastigotes. ExpParasitol. 2013 Mar;133(3):287-93.
- Michelini FM, Zorrilla P, Robello C, Alché LE. Immunomodulatory activity ofan anti-HSV-1 synthetic stigmastane analog. Bioorg Med Chem. 2013 Jan15;21(2):560-8.
2012
- Coitinho C, Greif G, Robello C, van Ingen J, Rivas C. Identification of Mycobacterium tuberculosis complex by polymerase chain reaction of Exact TandemRepeat-D fragment from mycobacterial cultures. Int J Mycobacteriol. 2012Sep;1(3):146-8.
- Pizzo, C., Faral-Tello, P., Salinas, G., Fló, M., Robello, C., Wipf, P., Mahler, G. Selenosemicarbazones as potent cruzipain inhibitors and their antiparasitic properties against Trypanosoma cruzi. MedChemCommun, 2012, 3, 362. DOI: 10.1039/c2md00283c
- Greif G, Coitinho C, Rivas C, van Ingen J, Robello C. Molecular analysis of isoniazid-resistant Mycobacterium tuberculosis isolates in Uruguay. Int J Tuberc Lung Dis. 2012 Jul;16(7):947-9.
- García G, Libisch G, Trujillo-Cenóz O, Robello C, Russo RE. Modulation of gene expression during early stages of reconnection of the turtle spinal cord. J Neurochem. 2012 Jun;121(6):996-1006.
- Peloso EF, Gonçalves CC, Silva TM, Ribeiro LH, Piñeyro MD, Robello C, GadelhaFR. Tryparedoxin peroxidases and superoxide dismutases expression as well as ROS release are related to Trypanosoma cruzi epimastigotes growth phases. ArchBiochem Biophys. 2012 Apr 15;520(2):117-22.
- Chiribao ML, Libisch MG, Osinaga E, Parodi-Talice A, Robello C. Cloning, localization and differential expression of the Trypanosoma cruzi TcOGNT-2 glycosyl transferase. Gene. 2012 May 1;498(2):147-54.
- Gascue C, Tan PL, Cardenas-Rodriguez M, Libisch G, Fernandez-Calero T, LiuYP, Astrada S, Robello C, Naya H, Katsanis N, Badano JL. Direct role of Bardet-Biedl syndrome proteins in transcriptional regulation. J Cell Sci. 2012Jan 15;125(Pt 2):362-75.
- Peñagaricano F, Zorrilla P, Naya H, Robello C, Urioste JI. Gene expression analysis identifies new candidate genes associated with the development of black skin spots in Corriedale sheep. J Appl Genet. 2012 Feb;53(1):99-106.
2011
- Peloso Ede F, Vitor SC, Ribeiro LH, Piñeyro MD, Robello C, Gadelha FR. Role of Trypanosoma cruzi peroxiredoxins in mitochondrial bioenergetics. J BioenergBiomembr. 2011 Aug;43(4):419-24.
- Piñeyro MD, Parodi-Talice A, Portela M, Arias DG, Guerrero SA, Robello C. Molecular characterization and interactome analysis of Trypanosoma cruzi tryparedoxin 1. J Proteomics. 2011 Aug 24;74(9):1683-92.
- Schijman AG, Bisio M, Orellana L, Sued M, Duffy T, Mejia Jaramillo AM, CuraC, Auter F, Veron V, Qvarnstrom Y, Deborggraeve S, Hijar G, Zulantay I, LuceroRH, Velazquez E, Tellez T, Sanchez Leon Z, Galvão L, Nolder D, Monje Rumi M, LeviJE, Ramirez JD, Zorrilla P, Flores M, Jercic MI, Crisante G, Añez N, De Castro AM, Gonzalez CI, Acosta Viana K, Yachelini P, Torrico F, Robello C, Diosque P,Triana Chavez O, Aznar C, Russomando G, Büscher P, Assal A, Guhl F, Sosa EstaniS, DaSilva A, Britto C, Luquetti A, Ladzins J. International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients.PLoS Negl Trop Dis. 2011 Jan 11;5(1):e931.
- Piñeyro MD, Arcari T, Robello C, Radi R, Trujillo M. Tryparedoxin peroxidases from Trypanosoma cruzi: high efficiency in the catalytic elimination of hydrogen peroxide and peroxynitrite. Arch Biochem Biophys. 2011 Mar 15;507(2):287-95.
2010
- Dujardin JC, Herrera S, do Rosario V, Arevalo J, Boelaert M, Carrasco HJ, Correa-Oliveira R, Garcia L, Gotuzzo E, Gyorkos TW, Kalergis AM, Kouri G, LarragaV, Lutumba P, Macias Garcia MA, Manrique-Saide PC, Modabber F, Nieto A, Pluschke G, Robello C, Rojas de Arias A, Rumbo M, Santos Preciado JI, Sundar S, Torres J, Torrico F, Van der Stuyft P, Victoir K, Olesen OF. Research priorities for neglected infectious diseases in Latin America and the Caribbean region. PLoSNegl Trop Dis. 2010 Oct 26;4(10):e780.