A5046956378- Animal Genetics and Reproduction
- CRISPR and Genetic Engineering
- Reproductive Biology and Fertility
- Pluripotent Stem Cells Research
- Sperm and Testicular Function
- RNA Interference and Gene Delivery
- Renal and related cancers
- Virus-based gene therapy research
- Viral Infectious Diseases and Gene Expression in Insects
- Reproductive Physiology in Livestock
- Prion Diseases and Protein Misfolding
- Genetic and Clinical Aspects of Sex Determination and Chromosomal Abnormalities
- Transgenic Plants and Applications
- Prenatal Screening and Diagnostics
- Assisted Reproductive Technology and Twin Pregnancy
- Advanced biosensing and bioanalysis techniques
- Reproductive Health and Technologies
University of Buenos Aires
2012-2022
Hospital de Clínicas "José de San Martín"
2022
Consejo Nacional de Investigaciones Científicas y Técnicas
2010-2020
In this work, Dehydroleucodine (DhL) was evaluated as a chemical activator of bovine oocytes and somatic cell nuclear transfer (SCNT) reconstituted embryos. Oocytes were activated with 5 microM Ionomycin (Io) exposed for 3 h to 1 or DhL alone (Io-Dhl1 Io-DhL5) combined Cytochalasin B (Io-DhL1/CB; Io-DhL5/CB). Control groups Io (Io), followed by 1.9 mM 6-Dimethylaminopurine (Io-6DMAP), embryos produced in vitro fertilization (IVF). Pronuclear formation development blastocysts evaluated....
Transgenic domestic animals represent an alternative to bioreactors for large-scale production of biopharmaceuticals and could also provide more accurate biomedical models than rodents. However, their generation remains inefficient. Recently, DNA transposons allowed improved transgenesis efficiencies in mice pigs. In this work, Tn5 Sleeping Beauty (SB) transposon systems were evaluated by simple cytoplasmic injection livestock zygotes. the case Tn5, transposome complex nucleic acid protein...
Summary The present study investigated the effects of crotamine, a cell-penetrating peptide from rattlesnake venom, at different exposure times and concentrations, on both developmental competence gene expression ( ATP1A1, AQP3, GLUT1 GLUT3 ) in vitro fertilized (IVF) bovine embryos. In Experiment 1, presumptive zygotes were exposed to 0.1 μM crotamine for 6, 12 or 24 h control groups (vehicle IVF) included. 2, 0 (vehicle), 0.1, 1 10 h. Additionally, visualize uptake, embryos rhodamine...
CRISPR-mediated transcriptional activation, also known as CRISPR-on, has proven efficient for activation of individual or multiple endogenous gene expression in cultured cells from several species. However, the potential CRISPR-on technology preimplantation mammalian embryos remains to be explored. Here, we report first time successful modulation bovine by using system. As a proof principle, targeted promoter region either SMARCA4 TFAP2C genes, transcription factors implicated trophoblast...
Microcell-mediated chromosome transfer (MMCT) was developed to introduce a low number of chromosomes into host cell. We have designed novel technique combining part MMCT with somatic cell nuclear transfer, which consists injecting micronucleus an enucleated oocyte, and inducing its cellular machinery replicate such micronucleus. It would allow the isolation manipulation single or chromosomes.Micronuclei from adult bovine fibroblasts were produced by incubation in 0.05 μg/ml demecolcine for...
Haploid activation of bovine oocytes after ICSI is a routine procedure. However, embryos frequently contain an abnormal chromosome set as result the drugs employed. We compared efficiency ionomycin (Io) followed by roscovitine (ROSC), cycloheximide (CHX), ethanol, and dehydroleucodine (DhL) to induce haploid parthenogenetic in bovine. Pronuclear (PN) formation, second polar body (2PB) extrusion, embryo development, ploidy blastocysts were evaluated. To this aim, COC aspirated from...
Haploid activation of bovine oocytes is important for reproductive technologies such as intracytoplasmic sperm injection (ICSI) or somatic cell nuclear transfer (SCNT). Nevertheless, it still a highly inefficient procedure. The aim this work was to combine different drugs, known have targets along the cascade, find more effective protocol. Cumulus-oocyte complexes (COC) were aspirated from slaughtered ovaries and in vitro-matured (IVM) 22 h. Oocytes activated with 5 µM ionomycin (IO) 4 min...
In bovine, the intracytoplasmic sperm injection (ICSI) technique remains inefficient probably because of low levels male decondensation. species with frequent fertilization failure, high cysteamine (Cys) concentration during in vitro maturation (IVM) has been used to improve IVF. Cysteamine, a precursor glutathione, plays critical role on The aim this work was ICSI efficiency bovine by (1) increasing endogenous glutathione from oocytes using Cys IVM; and (2) incubating cumulus-oocyte...
Techniques available for production of transgenic animals remain inefficient. Recently an effective transgenesis system using I-SceI meganuclease (MegaN) has been described in Xenopus. It consists MegaN incubation with a pIS-plasmid carrying recognition sites-, followed by injection into fertilized egg. Here, we evaluated the combined new approaches developed our group. We compared (1) three pIS concentrations incubated or not MegaN; (2) oocytes at germinal vesicle stage (GV) presumptive...
Intracytoplasmic sperm injection (ICSI) mediated transgenesis is an effective tool for transgenic animal production. However, ICSI in cattle remains inefficient. In this work, we assayed approaches to improve egfp expressing blastocysts production by ICSI: the pretreatment with heparin and l-glutathione (Hep-GSH), use of sex-sorted (SS), refrozen/thawing SS sperm, combination these. Quality was analysed studying expression 4 genes, rates DNA fragmentation. Cumulus-oocyte complexes from...
Current techniques for the production of transgenic domestic animals remain inefficient. Only recently, DNA transposons resulted in improved efficiencies mouse and pig transgenesis. In this work, we evaluated Tn5 Sleeping Beauty systems transgenesis bovine ovine species. First, both transposon were assessed vitro embryos employing carrying fluorescent reporter genes. vitro-produced zygotes microinjected with either 1) a complex Tn5:egfp (20 ng μL–1) (protein: transgene mosaic ends recognised...
The rapid introduction of engineered nucleases technologies, such as zinc finger (ZFN), transcription activator-like effector (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR), provides new opportunities for editing genes in a targeted rather simple fashion. Few reports are available regarding CRISPR efficiency domestic species. Here, the CRISPR/Cas9 system was employed to develop knockout knock-in alleles bovine PRNP gene, responsible spongiform encephalopathy...
Reduced glutathione (GSH) is an endogenous disulfide bond reducer present in mammalian oocytes. It plays a critical role sperm decondensation following fertilization, disrupting the protamine bonds that sustain hypercondensed state of DNA. However, reduction needs to be followed by removal achieve male pronuclear formation. In humans, heparan sulfate (HS) has been shown exert this (Romanato et al. 2008 Hum. Reprod. 23, 1145–1450). Although there are no reports bovine, we recently...
Cloning of domestic species by somatic cell nuclear transfer (SCNT) continues to be inefficient, probably due an incomplete reprogramming the reconstituted embryo. The ability embryonic cytoplasm support fluctuates within cycle (Egli et al. 2007 Nature 447, 679–85). In this context, we compared development capability and second polar body (2PB) extrusion embryos produced metaphase (M) cells, in comparison with G0/G1 which are commonly used as donors. Because M cells have 2 sets chromosomes...
Microcell-mediated chromosome transfer was developed to introduce a low number of chromosomes into host cell (Fournier and Ruddle 1977 PNAS 74, 319–323). As first approach individual manipulation, we designed new technique that consists injecting micronucleus an enucleated oocyte replicate chromosomes. Additionally, studied the capability such micronuclei be marked with transgene. To this aim, from adult bovine fibroblasts were produced by incubation in 0.05 μg mL–1 demecolcine for 48 h...