A5048938769- Animal Genetics and Reproduction
- Reproductive Biology and Fertility
- CRISPR and Genetic Engineering
- Sperm and Testicular Function
- Pluripotent Stem Cells Research
- Renal and related cancers
- Ovarian function and disorders
- Neuroscience and Neuropharmacology Research
- Genetics and Neurodevelopmental Disorders
- Reproductive System and Pregnancy
- Epigenetics and DNA Methylation
- Virus-based gene therapy research
- Urological Disorders and Treatments
- Reproductive biology and impacts on aquatic species
- Molecular Biology Techniques and Applications
- Transgenic Plants and Applications
- Renin-Angiotensin System Studies
- Metabolomics and Mass Spectrometry Studies
- RNA Research and Splicing
- bioluminescence and chemiluminescence research
- Cellular transport and secretion
- Tendon Structure and Treatment
- Genomics and Chromatin Dynamics
- Hormonal Regulation and Hypertension
- MicroRNA in disease regulation
Iwate University
2017-2024
Kyoto University
2011-2022
The University of Tokyo
2015
Hamamatsu University School of Medicine
2015
Central Institute for Experimental Animals
2015
Weatherford College
2014
Kumamoto University
2003-2011
University of Hawaii System
2005-2006
University of Hawaiʻi at Mānoa
2003
Yamagata University
2001
Abstract The CRISPR-Cas system is a powerful tool for generating genetically modified animals; however, targeted knock-in (KI) via homologous recombination remains difficult in zygotes. Here we show efficient gene KI rats by combining with single-stranded oligodeoxynucleotides (ssODNs). First, 1-kb ssODN co-injected guide RNA (gRNA) and Cas9 messenger produce GFP-KI at the rat Thy1 locus. Then, two gRNAs 80-bp ssODNs direct integration of 5.5-kb CAG-GFP vector into Rosa26 locus...
Transcription activator-like effector (TALE) nuclease (TALEN) is a site-specific nuclease, which can be freely designed and easily constructed. Numerous methods of constructing TALENs harboring different TALE scaffolds repeat variants have recently been reported. However, the functionalities structurally not yet compared. Here, we report on functional differences among several types targeting same loci. Using HEK293T cell-based single-strand annealing Cel-I assays, found that with...
Engineered endonucleases, such as zinc-finger nucleases (ZFNs), transcription activator-like effector (TALENs), and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system, provide a powerful approach for genome editing in animals. However, microinjection of endonucleases into embryos requires high skill level, is time consuming, may cause damage to embryos. Here, we demonstrate that electroporation endonuclease mRNAs intact can induce at targeted...
Abstract The bacterial CRISPR/Cas system has proven to be an efficient gene-targeting tool in various organisms. Here we employ for accurate and genome editing rats. synthetic chimeric guide RNAs (gRNAs) discriminate a single-nucleotide polymorphism (SNP) difference rat embryonic fibroblasts, allowing allele-specific of the dominant phenotype (F344 × DA)F1 hybrid embryos. Interestingly, targeted allele, initially assessed by gRNA, is repaired interallelic gene conversion between homologous...
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system is a powerful tool for genome editing in animals. Recently, new technology has been developed to genetically modify animals without using highly skilled techniques, such as pronuclear microinjection of endonucleases. Technique animal knockout by electroporation (TAKE) method simple and effective that produces rats introducing endonuclease mRNAs into intact embryos electroporation. Using TAKE...
DEP-domain containing 5 (DEPDC5), encoding a repressor of the mechanistic target rapamycin complex 1 (mTORC1) signaling pathway, has recently emerged as major gene mutated in familial focal epilepsies and cortical dysplasia. Here we established global knockout rat using TALEN technology to investigate vivo impact Depdc5-deficiency. Homozygous Depdc5(-/-) embryos died from embryonic day 14.5 due growth delay. Constitutive mTORC1 hyperactivation was evidenced brains cultured fibroblasts...
Embryonic appendicular structures, such as the limb buds and developing external genitalia, are suitable models with which to analyze reciprocal interactions of growth factors in regulation outgrowth. Although several studies have evaluated individual functions different growth, coordinated function integration input from multiple signaling cascades is poorly understood. We demonstrate that a novel cascade governs formation embryonic genitalia [genital tubercle (GT)]. show dosage Shh signal...
TAL Effector Nucleases (TALENs) are versatile tools for targeted gene editing in various species. However, their efficiency is still insufficient, especially mammalian embryos. Here, we showed that combined expression of Exonuclease 1 (Exo1) with engineered site-specific TALENs provided highly efficient disruption the endogenous rat fibroblast cells. A similar increased up to ~30% Exo1 was also observed fertilized eggs, and production knockout rats albino (Tyr) gene. These findings...
The transcription factor Nrf2 (NF-E2-related-factor 2) regulates a battery of antioxidative stress-response genes and detoxication genes, knockout lines mice have been contributing critically to the clarification roles that plays for cell protection. However, there are apparent limitations in use mouse models. For instance, rats exhibit more suitable features toxicological or physiological examinations than mice. In this study, we generated 2 by using genome editing technology; 1 line...
The epididymis is a male accessory organ and functions for sperm maturation storage under the control of androgen. development also androgen dependent. Wolffian duct (WD), anlagen epididymis, formed in both female embryos; however, it stabilized only embryos by testicular Androgen drives subsequent differentiation WD into epididymis. Although essential roles masculinization epididymal function have been established, little known about cellular events regulated precisely signaling during...
The protective effect of aliskiren, a direct renin inhibitor, against hypertensive cardiovascular and renal injury remains to be defined. This study was undertaken examine the effects combination aliskiren valsartan, an angiotensin receptor blocker, injury. Endothelial NO synthase–deficient mice, subjected cuff femoral artery, were divided into 5 groups treated with following: (1) vehicle; (2) (25 mg/kg per day); (3) valsartan (8 (4) combined (12.5 day) (4 (5) hydralazine (10 for 4 weeks....
Mouse spermatozoa from the caudae epididymides could be freeze-dried without losing their ability to support normal development. Immature testes, in contrast, were damaged by freeze-drying. However, immature became resistant freeze-drying after treatment with diamide, which oxidizes free -SH groups. Conversely, epididymal if first treated dithiothreitol (DTT), reduces -SS- bonds. The potential for damage seems likely relate status of sperm proteins, particular its protamines.
Significance Genomically humanized animals overcoming species differences are invaluable for biomedical research. Although rats would be preferred over mice several applications, generation of a model is restricted to due the difficulty complex genetic manipulations in rats. In this study, we successfully generated with megabase-sized gene clusters via combination chromosome transfer using mouse artificial vector and genome editing technologies. UGT2 CYP3A transchromosomic described paper,...
The number of endangered avian-related species increase in Japan recently. application new technologies, such as induced pluripotent stem cells (iPSCs), may contribute to the recovery decreasing numbers animals and conservation genetic resources. We established novel iPSCs from three avian (Okinawa rail, Japanese ptarmigan, Blakiston's fish owl) with seven reprogramming factors (M3O, Sox2, Klf4, c-Myc, Nanog, Lin28, Klf2). are pluripotency markers express pluripotency-related genes...
Freeze-drying sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying that it can be stored at 4 °C and transported room temperature. Although the successful reported in number animals, possibility long-term using this not yet studied.Offspring were obtained from oocytes fertilized with rat epididymal freeze-dried solution containing 10 mM Tris 1 EDTA adjusted to pH 8.0. Tolerance testicular was increased by...
Sperm preservation is a useful technique for the maintenance of biological resources in experimental and domestic animals, wild animals. A new method has been developed that enables sperm to be stored long time refrigerator at 4°C. are freeze-dried solution containing 10 mM Tris 1 EDTA. Using this method, liquid nitrogen not required storage transportation sperm. We demonstrate chimpanzee, giraffe, jaguar, weasel long-haired rat remain viable after freeze-drying. In all species, pronuclei...
Background and Purpose- oxLDL (oxidized low-density lipoprotein) has been known for its potential to induce endothelial dysfunction used as a major serological marker of oxidative stress. Recently, LOX-1 (lectin-like oxidized lipoprotein receptor-1), lectin-like receptor oxLDL, attracted attention in studies neuronal apoptosis stroke. We aim investigate the impact LOX-1-deficiency on spontaneous hypertension-related brain damage present study. Methods- generated deficient strain genetic...
The protein CHD1 is a member of the family ATPase-dependent chromatin remodeling factors. CHD1, which recognizes trimethylated histone H3 lysine 4, has been implicated in transcriptional activation organisms ranging from yeast to humans. It required for pre-mRNA maturation, maintenance mouse embryonic stem cell pluripotency and rapid growth epiblast. However, function(s) preimplantation embryos not yet examined. Here, we show that loss function led lethality after implantation. In Chd1 was...
SET and MYND domain-containing protein 3 ( Smyd3 ) is a histone H3 lysine 4 (H3K4) di- tri-methyltransferase that forms transcriptional complex with RNA polymerase II activates the transcription of oncogenes cell cycle genes in human cancer cells. However, study mammalian early embryonic development has not yet been addressed. In present study, we investigated expression pattern mouse preimplantation embryos effects interference (RNAi)-mediated repression on embryos. We showed mRNA levels...