A5067166467- CRISPR and Genetic Engineering
- Animal Genetics and Reproduction
- Reproductive Biology and Fertility
- Pluripotent Stem Cells Research
- Sperm and Testicular Function
- Genetic and phenotypic traits in livestock
- Viral Infectious Diseases and Gene Expression in Insects
- Reproductive Health and Technologies
- Virus-based gene therapy research
- Selenium in Biological Systems
- Aldose Reductase and Taurine
- RNA regulation and disease
- Bee Products Chemical Analysis
- Ovarian function and disorders
- CAR-T cell therapy research
- Viral Infections and Immunology Research
- Effects of Environmental Stressors on Livestock
- Sesame and Sesamin Research
- Reproductive Physiology in Livestock
- Cancer-related molecular mechanisms research
- Cannabis and Cannabinoid Research
- Animal Behavior and Welfare Studies
- Tissue Engineering and Regenerative Medicine
- Hemoglobinopathies and Related Disorders
- Neonatal Health and Biochemistry
Tokushima University
2023-2025
Yamagata Prefectural Central Hospital
2022
ABSTRACT Pig genome editing using the oviductal nucleic acid delivery (GONAD) method with electroporation would allow efficient obtention of genetically modified pigs. However, oocytes and zygotes at various stages after ovulation must be targeted, cumulus cell attachment mosaic mutations are major obstacles. Therefore, we investigated whether two parameters (electroporation timing attachment) influence effectiveness multiplex by in porcine or zygotes. Three gRNAs targeting either GGTA1 ,...
Although yaks and cattle belong to the same Bovinae subfamily have number of chromosomes, hybrid males are sterile because inactivation or abnormality gene expression related production healthy normal sperm. Recently, analysis not only in testis but also epididymis has offered hints about mechanism infertility, supports maturation sperm acquiring capacity fertilisation. Sperm processes been thought be androgen-dependent, androgen receptor (AR) can activated by dihydrotestosterone converted...
Abstract This study examined the effects of ergothioneine (EGT) supplementation as an antioxidant on quality boar spermatozoa when using liquid and frozen preservation methods. In first experiment, semen was preserved in extender supplemented with 0, 50, 100 200 µM EGT, at 15 °C, part samples for one another three weeks. comparison control (without EGT), EGT significantly increased percentage total motility that were a both weeks ( P < 0.05). did not affect spermatozoa, irrespective...
Abstract The transfection efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)/Cas ribonucleoprotein complexes was compared using three nonviral vector reagents: nonliposomal polymeric (TransIT‐X2), lipid nanoparticle delivery (CRISPRMAX), and peptide (ProteoCarry) systems. Porcine zona pellucida‐free zygotes embryos were incubated for 5 h with CRISPR‐associated protein 9 (Cas9), guide RNA (gRNA) targeting GGTA1, one the reagents. In Experiment 1, optimization...
We evaluated the quality and fertilizing ability of frozen-thawed porcine sperm that were selected using a commercially available device (MIGLIS, Menicon Life Science) consisting three parts: an outer lid, inner tube. Firstly, to determine adequate concentration caffeine for separation, incubated with different concentrations (0, 1, 2.5, 5, 10 mM) in MIGLIS device. To appropriate incubation time separating device, 2.5 mM 10, 15, or 20 min. evaluate fertilization embryo development oocytes...
CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9) technology is growing rapidly and has been greatly influencing the efficiency effectiveness of genetic modifications in different applications. One aspect research gaining importance development system introduction CRISPR materials into target organisms. Although we previously demonstrated efficacy electroporation- lipofection-mediated gene disruption porcine zygotes, still believe that this...
Dairy farmers and beef cattle breeders aim for one calf per year to optimize breeding efficiency, relying on artificial insemination of both dairy cows. Accurate estrus detection timely are vital improving conception rates. However, recent challenges such as operational expansion, increased livestock numbers, heightened milk production have complicated these processes. We developed an intelligence (AI)-based pregnancy probability diagnostic tool predict the optimal timing insemination. This...
The antioxidant effects of ergothioneine (EGT) on
CRISPR/Cas9-based multiplex genome editing via electroporation is relatively efficient; however, lipofection versatile because of its ease use and low cost. Here, we aimed to determine the efficiency in using growth hormone receptor (GHR) glycoprotein alpha-galactosyltransferase 1 (GGTA1)-targeting guide RNAs (gRNAs) pig zygotes. Zona pellucida-free zygotes were collected 10 h after vitro fertilization incubated with Cas9, gRNAs, Lipofectamine 2000 (LP2000) for 5 h. In Experiment 1,...
Mosaicism, which is characterized by the presence of wild-type and more than one mutant allele, poses a serious problem in zygotic gene modification through clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 system. Therefore, we used pig embryos to compare editing efficiencies achieved combining electroporation lipofection using different aminopeptidase N (APN)-targeting guide RNA (gRNA) sequences. Six gRNAs (gRNA1-6) with target sequences were...
BACKGROUND: Cryopreservation of bovine zygotes allows for a flexible schedule genome editing via electroporation. However, vitrification-induced cell membrane damage may not only affect embryonic development but also mutation. OBJECTIVE: To investigate the effects vitrification before and after electroporation treatments on mutation presumptive zygotes. MATERIALS AND METHODS: In vitro-derived were electroporated with CRISPR/Cas9 system immediately (Vitrified-EP) or 2 h incubation...
Genetic mosaicism is considered one of the main limitations electroporation method used to transfer CRISPR-Cas9/guide RNA (gRNA) into porcine zygotes. We hypothesized that fertilization oocytes with sperm from gene-deficient boars, in combination (EP) target same region gene subsequent zygotes, would increase modification efficiency. As myostatin (MSTN) and α1,3-galactosyltransferase (GGTA1) have beneficial effects on agricultural production xenotransplantation, respectively, we these two...
Pigs rarely develop cancer; however, tumour protein p53 (TP53)-modified pigs may have an increased incidence of cancer. In this study, two with mosaic mutations induced by gene editing were compared to determine the role wild-type TP53 sequence in tumorigenesis and speculate how amino acid changes sequences are related tumorigenesis. The pig without tumours had a 1-bp deletion that resulted premature stop codon. contrast, nephroblastoma 6- 7-bp deletions sequence, resulting absence acids...
Abstract. This study investigated the relationship between oxidation and antioxidation parameters before superovulation (SOV) treatment embryo recovery in donor cows. The relative redox status of 61 cows was evaluated based on median values diacron-reactive oxygen metabolites (d-ROMs) biological antioxidant potential (BAP) measurements (d-ROMs 100 U.CARR; BAP 2413 µmol L−1) SOV treatment. Following this, animals were divided into four groups: with low d-ROMs but high assigned to “LH” group...
Abstract Sterilization of the culture medium using ultraviolet (UV)‐C reduces potential adverse effects microorganisms and allows for long‐term use. In present study, we investigated a directly irradiated with UV‐C prior to in vitro on development quality porcine vitro‐fertilized embryos free amino acid composition media. The media (porcine zygote [PZM‐5] blastocyst [PBM]) were at 228 260 nm 1 3 days, respectively. Next, 3, 7, or 14 days. After fertilization, cultured UV‐C‐irradiated 7 Free...