A5065737224- Reproductive Biology and Fertility
- CRISPR and Genetic Engineering
- Animal Genetics and Reproduction
- Sperm and Testicular Function
- Pluripotent Stem Cells Research
- Reproductive Physiology in Livestock
- Assisted Reproductive Technology and Twin Pregnancy
- Xenotransplantation and immune response
- Pregnancy and preeclampsia studies
- Virus-based gene therapy research
- Animal Behavior and Welfare Studies
- Reproductive Health and Technologies
- Genetic and phenotypic traits in livestock
- MicroRNA in disease regulation
- Innovation and Socioeconomic Development
- Urological Disorders and Treatments
- Agriculture and Biological Studies
- Reproductive biology and impacts on aquatic species
- Genetic Mapping and Diversity in Plants and Animals
- Extracellular vesicles in disease
- Gestational Diabetes Research and Management
- Calcium signaling and nucleotide metabolism
- Neonatal Health and Biochemistry
- Circular RNAs in diseases
- Reproductive System and Pregnancy
Centro de Investigacion Principe Felipe
2024-2025
Universidad de Murcia
2019-2024
Instituto Murciano de Investigación Biosanitaria
2019-2024
University of California, Davis
2020
Genetically modified pigs play a critical role in mimicking human diseases, xenotransplantation, and the development of resistant to viral diseases. The use programmable endonucleases, including CRISPR/Cas9 system, has revolutionized generation genetically pigs. This study evaluates efficiency electroporation oocytes prior fertilization generating edited gene embryos for different models. For single editing, phospholipase C zeta (PLC ζ) fused sarcoma (FUS) genes were used, concentration...
Studies of knockout (KO) mice with defects in the endolysosomal two-pore channels (TPCs) have shown TPCs to be involved pathophysiological processes, including heart and muscle function, metabolism, immunity, cancer, viral infection. With objective studying TPC2's roles for first time a large, more humanlike animal model, TPC2 KO pigs were produced using CRISPR-Cas9. A major problem CRISPR-Cas9 edit embryos is mosaicism; thus, we studied effect microinjection timing on mosaicism. Mosaicism...
The generation of genetically modified pigs has an important impact thanks its applications in basic research, biomedicine, and meat production. Cloning was the first technique used for this production, although easier cheaper methods were developed, such as microinjection, electroporation, or lipofection oocytes zygotes. In study, we analyzed production embryos via zona-pellucida-intact using LipofectamineTM CRISPRMAXTM Cas9 comparison with electroporation method. Two factors evaluated: (i)...
Context Extracellular vesicles (EVs) derived from the oviductal fluid (oEVs) play a critical role in various reproductive processes, including sperm capacitation, fertilisation, and early embryo development. Aims To characterise porcine oEVs (poEVs) different stages of estrous cycle (late follicular, LF; luteal, EL; mid ML; late LL) investigate their impact on functionality. Methods poEVs were isolated, characterised, labelled to assess binding boar spermatozoa. The effects motility,...
The evolution of some fetal growth indices and arterial blood flow parameters in the umbilical cord embryos fetuses primiparous pregnant goats Murciano-Granadina breed were analyzed by ultrasonography. Weekly ultrasonographic sessions took place from 18- to 125-days post-breeding. Fetal measures carried out ultrasound B-mode. This mode was used take a series measurements embryo/fetus throughout pregnancy: crown-rump length (CRL, 24-days post-mating -dpm- 61 dpm), trunk diameter (TD, 24–34...
Abstract Background In vitro embryo production (IVP) and transfer (ET) are two very common assisted reproductive technologies (ART) in human cattle. However, pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to low efficiency IVP plus difficulty performing ET long contorted uterus sow. addition, potential impact these ART on health offspring unknown. We investigated here if a modified system, with natural fluids (RF) as supplements...
Mosaicism is the most important limitation for one-step gene editing in embryos by CRISPR/Cas9 because cuts and repairs sometimes take place after first DNA replication of zygote. To try to minimize risk mosaicism, this study a reversible inhibitor was used release cell. There no previous information on use aphidicolin porcine embryos, so inhibition effect embryo development different concentrations drug evaluated. The incubation with tested at delivery methodologies. As result, observed, it...
Limb girdle muscular dystrophy type R1 (LGMDR1) is an autosomal recessive myopathy described in humans resulting from a deficiency of calpain-3 protein (CAPN3). This disease lacks effective treatment and appropriate model, so the generation KO pigs by CRISPR-Cas9 offers way to better understand ethology develop novel therapies. Microinjection main method for gene editing porcine embryo, but electroporation, which allows handling more embryos faster easier, has also recently been reported....
Assisted reproductive technologies play a major role in the cattle industry. An increase use of vitro -derived embryos is currently being seen around globe. But efficiency and quality are substandard when compared to vivo production. Different protocols have been designed overcome this issue, one those fluids as supplementation embryo culture media. In study, calves produced with added their production protocol were followed for first year life pairwise control, by artificial insemination...
More suitable and efficient methods to protect gametes from external harmful effects during in vitro handling can be achieved by adding preovulatory porcine oviductal fluid (pOF) culture media. The objective of this study was assess the swim-up procedure’s suitability as a sperm selection method using medium supplemented with 1mg/mL BSA, 1% pOF (v/v), v/v plus 5mg/mL BSA. After selection, various parameters were studied, such recovery rate, morphology, motility (by CASA), vitality, acrosome...
Over the last 50 years, fertility of high-producing dairy cows has declined due to intensity genetic selection. To overcome this obstacle, assisted reproductive techniques have been used. However, in vitro embryo production is a relatively inefficient process, with maximum blastocyst yield rates around 35%. problem, it was hypothesised that by adding endometrial organoid secretions culture medium, embryos produced would be exposed more physiological environment and technique increase. For...
Introduction: CRISPR/Cas9-mediated editing of embryos through microinjection is limited in efficiency because high rates mosaicism and off-target mutations. Electroporation has been proposed as an easier faster procedure compared to single embryo injection. Methods: Using a synthetic guide-RNA targeting porcine NGN3, we optimized the electroporation conditions effectively introduce CRISPR/Cas9 zygotes evaluate its effects on off-targets. Results: A numerical increase mutation was observed...
High-grade serous ovarian cancer (HGSOC) is the most lethal form of gynecologic cancer, with limited treatment options and a poor prognosis. Epigenetic factors, such as microRNAs (miRNAs) DNA methylation, play pivotal roles in progression, yet their specific contributions to HGSOC remain insufficiently understood. In this study, we performed comprehensive high-throughput analyses identify dysregulated miRNAs investigate epigenetic regulation. Analysis tissue samples from advanced-stage...
Each year, tens of thousands people worldwide die end-stage organ failure due to the limited availability organs for use in transplantation. To meet this clinical demand, one last frontiers regenerative medicine is generation humanized pigs from pluripotent stem cells (PSCs) via blastocyst complementation. For this, organ-disabled pig models are needed. As endothelial (ECs) play a critical role xenotransplantation rejection every organ, we aimed produce hematoendothelial-disabled embryos...
Assisted reproductive technologies (ART) are being extensively used to produce cattle offspring. However, as shown by Siqueira et al. (2017 J. Dairy Sci. 100, 5899-5908; https://doi.org/10.3168/jds.2016-12539), phenotypical and performance differences between cows derived from distinct ART can be found at different stages of development. Thus, in an attempt mimic the natural environment, fluids (oviductal (SOF) uterine fluids) were added supplementation embryo culture media. Our hypothesis...
Abstract Background : In vitro embryo production (IVP) and transfer (ET) are two very common assisted reproductive technologies (ART) in human cattle. However, pig, the combination of either procedures, or even their use separately, is still considered suboptimal due to low efficiency IVP plus difficulty performing ET long contorted uterus sow. addition, potential impact these ART on health offspring unknown. We investigated here if a modified system, with natural fluids (RF) as supplements...