1,500 results match your criteria generated egfp

Dual role of miR-1 in the development and function of sinoatrial cells.

J Mol Cell Cardiol 2021 May 5. Epub 2021 May 5.

Department of Biosciences, Università degli Studi di Milano, Milan, Italy. Electronic address:

miR-1, the most abundant miRNA in the heart, modulates expression of several transcription factors and ion channels. Conditions affecting the heart rate, such as endurance training and cardiac diseases, show a concomitant miR-1 up- or down-regulation. Here, we investigated the role of miR-1 overexpression in the development and function of sinoatrial (SAN) cells using murine embryonic stem cells (mESC). Read More

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Neural stem cells in the adult olfactory bulb core generate mature neurons in vivo.

Stem Cells 2021 May 8. Epub 2021 May 8.

Instituto Cajal-Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain.

Although previous studies suggest that neural stem cells (NSCs) exist in the adult olfactory bulb (OB), their location, identity and capacity to generate mature neurons in vivo has been little explored. Here, we injected EGFP-expressing retroviral particles into the OB core of adult mice to label dividing cells and to track the differentiation/maturation of any neurons they might generate. EGFP labelled cells initially expressed adult NSC markers on days 1-3 post injection (dpi), including Nestin, GLAST, Sox2, Prominin-1 and GFAP. Read More

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Modulating innate immune activation states impacts the efficacy of specific Aβ immunotherapy.

Mol Neurodegener 2021 May 6;16(1):32. Epub 2021 May 6.

Department of Neuroscience and Neurology, Center for Translational Research in Neurodegenerative Disease, and McKnight Brain Institute, University of Florida, FL, 32611, Gainesville, USA.

Introduction: Passive immunotherapies targeting Aβ continue to be evaluated as Alzheimer's disease (AD) therapeutics, but there remains debate over the mechanisms by which these immunotherapies work. Besides the amount of preexisting Aβ deposition and the type of deposit (compact or diffuse), there is little data concerning what factors, independent of those intrinsic to the antibody, might influence efficacy. Here we (i) explored how constitutive priming of the underlying innate activation states by Il10 and Il6 might influence passive Aβ immunotherapy and (ii) evaluated transcriptomic data generated in the AMP-AD initiative to inform how these two cytokines and their receptors' mRNA levels are altered in human AD and an APP mouse model. Read More

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Fe(III)-complex mediated bacterial cell surface immobilization of eGFP and enzymes.

Chem Commun (Camb) 2021 May;57(36):4460-4463

Lehrstuhl für Biotechnologie, RWTH Aachen University, Worringerweg 3, Aachen 52074, Germany. and DWI - Leibniz Institut für Interaktive Materialien, Forckenbeckstraße 50, 52074, Aachen, Germany.

We report a facile and reversible method to immobilize a broad range of His6-tagged proteins on the E. coli cell surface through Fe(iii)-metal complexes. A His6-tagged eGFP and four His6-tagged enzymes were successfully immobilized on the cell surface. Read More

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iTIME.219: An Immortalized KSHV Infected Endothelial Cell Line Inducible by a KSHV-Specific Stimulus to Transition From Latency to Lytic Replication and Infectious Virus Release.

Front Cell Infect Microbiol 2021 14;11:654396. Epub 2021 Apr 14.

Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, United States.

Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) is the causative agent of Kaposi's sarcoma and two B cell lymphoproliferative disorders: primary effusion lymphoma and KSHV-associated multicentric Castleman's disease. These distinct pathologies involve different infected cell types. In Kaposi's sarcoma, the virus is harbored in spindle-like tumor cells of endothelial origin, in contrast with the two pathologies of B cells. Read More

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Generation of a Novel Transgenic Zebrafish for Studying Adipocyte Development and Metabolic Control.

Int J Mol Sci 2021 Apr 13;22(8). Epub 2021 Apr 13.

Hanbang Cardio-Renal Research Center, Wonkwang University, Iksan 54538, Korea.

Zebrafish have become a popular animal model for studying various biological processes and human diseases. The metabolic pathways and players conserved among zebrafish and mammals facilitate the use of zebrafish to understand the pathological mechanisms underlying various metabolic disorders in humans. Adipocytes play an important role in metabolic homeostasis, and zebrafish adipocytes have been characterized. Read More

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Pupae Efficiently Produce Recombinant AAV2/HBoV1 Vectors with a Nuclear Polyhedrosis Virus Expression System.

Viruses 2021 04 18;13(4). Epub 2021 Apr 18.

School of Life Sciences, Jiangsu University, Zhenjiang 212013, China.

Recombinant adeno-associated virus (AAV) vectors have broad application prospects in the field of gene therapy. The establishment of low-cost and large-scale manufacturing is now the general agenda for industry. The baculovirus-insect cell/larva expression system has great potential for these applications due to its scalability and predictable biosafety. Read More

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Reverse genetics systems for contemporary isolates of respiratory syncytial virus enable rapid evaluation of antibody escape mutants.

Proc Natl Acad Sci U S A 2021 Apr;118(14)

Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine Hannover, 30559 Hannover, Germany;

Human respiratory syncytial virus (RSV) is the leading cause of acute lower respiratory infection in children under 5 y of age. In the absence of a safe and effective vaccine and with limited options for therapeutic interventions, uncontrolled epidemics of RSV occur annually worldwide. Existing RSV reverse genetics systems have been predominantly based on older laboratory-adapted strains such as A2 or Long. Read More

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A Model System to Explore the Detection Limits of Antibody-Based Immuno-SPECT Imaging of Exclusively Intranuclear Epitopes.

J Nucl Med 2021 Mar 31. Epub 2021 Mar 31.

Medical Research Council Oxford Institute for Radiation Oncology, Department of Oncology, University of Oxford, United Kingdom.

Imaging of intranuclear epitopes using antibodies tagged to cell penetrating peptides (CPPs) has great potential given its versatility, specificity and sensitivity. However, this process is technically challenging because of the location of the target. Previous research has demonstrated a variety of intranuclear epitopes that can be targeted with antibody-based radioimmunoconjugates (RICs). Read More

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Prolonged activation of cytomegalovirus early gene e1-promoter exclusively in neurons during infection of the developing cerebrum.

Acta Neuropathol Commun 2021 03 9;9(1):39. Epub 2021 Mar 9.

Department of Regenerative & Infectious Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-Ku, Hamamatsu, 431-3192, Japan.

The brain is the major target of congenital cytomegalovirus (CMV) infection. It is possible that neuron disorder in the developing brain is a critical factor in the development of neuropsychiatric diseases in later life. Previous studies using mouse model of murine CMV (MCMV) infection demonstrated that the viral early antigen (E1 as a product of e1 gene) persists in the postnatal neurons of the hippocampus (HP) and cerebral cortex (CX) after the disappearance of lytic infection from non-neuronal cells in the periventricular (PV) region. Read More

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Adaptation of Gut Microbiome to Transgenic Pigs Secreting β-Glucanase, Xylanase, and Phytase.

Front Genet 2021 4;12:631071. Epub 2021 Mar 4.

National Engineering Research Center for Breeding Swine Industry, Wens Foodstuff Group Co., Ltd., Yunfu, China.

We previously generated transgenic pigs with enhanced growth rate and reduced nutrient loss. However, the composition of their gut microbiome is unknown. In this study, we successfully generated marker-free transgenic (MF-TG) pigs with high expression levels of microbial β-glucanase, xylanase, and phytase in the parotid gland. Read More

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Highly Efficient Genome Modification of Cultured Primordial Germ Cells with Lentiviral Vectors to Generate Transgenic Songbirds.

Stem Cell Reports 2021 Apr 18;16(4):784-796. Epub 2021 Mar 18.

Max Planck Institute for Ornithology, Eberhard Gwinner Strasse, 82319 Seewiesen, Germany.

The ability to genetically manipulate organisms has led to significant insights into functional genomics in many species. In birds, manipulation of the genome is hindered by the inaccessibility of the one-cell embryo. During embryonic development, avian primordial germ cells (PGCs) migrate through the bloodstream and reach the gonadal anlage, where they develop into mature germ cells. Read More

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Three-dimensional microscopy and image fusion reconstruction analysis of the thyroid gland during morphogenesis.

FEBS Open Bio 2021 May 1;11(5):1417-1427. Epub 2021 Apr 1.

Department of Molecular Diagnostics & Endocrinology, The Core Laboratory in Medical Center of Clinical Research, State Key Laboratory of Medical Genomics, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, China.

Thyroid dysgenesis (TD) is a major cause of primary congenital hypothyroidism; however, the molecular mechanism underlying this process is unclear. Current knowledge regarding the morphogenesis of the thyroid gland and vascular anomalies affecting thyroid development is limited. To monitor the early stages of thyroid gland development, we generated double transgenic zebrafish embryos Tg(tg:mCherry/flk1:EGFP). Read More

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Replication of heterologous glycoprotein-expressing chimeric recombinant snakehead rhabdoviruses (rSHRVs) and viral hemorrhagic septicemia viruses (rVHSVs) at different temperatures.

Virus Res 2021 May 13;297:198392. Epub 2021 Mar 13.

Department of Aquatic Life Medicine, Pukyong National University, Busan, 48513, South Korea. Electronic address:

Water temperature is an important environmental factor for the outbreaks of fish rhabdovirus diseases. In the present study, to know the role of piscine rhabdoviral glycoproteins in the determination of replication temperature, several chimeric snakehead rhabdoviruses (SHRVs) and viral hemorrhagic septicemia viruses (VHSVs) expressing heterologous glycoproteins (rSHRV-Gvhsv, SHRV expressing VHSV G protein; rSHRV-Gsvcv, SHRV expressing spring viremia of carp virus G protein; rVHSV-Gshrv, VHSV expressing SHRV G protein; rVHSV-Gsvcv, VHSV expressing SVCV G protein) were generated using reverse genetics, and their replication characteristics at different temperatures were investigated. Furthermore, based on SHRV minigenome containing a reporter gene, the role of VHSV N, P, and L proteins in the determination of VHSV's low-temperature replication was investigated. Read More

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Striated myosin heavy chain gene is a crucial regulator of larval myogenesis in the pacific oyster Crassostrea gigas.

Int J Biol Macromol 2021 May 6;179:388-397. Epub 2021 Mar 6.

Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China; Laboratory of Tropical Marine Germplasm Resources and Breeding Engineering, Sanya Oceanographic Institution, Ocean University of China, Sanya 572000, China. Electronic address:

Pacific oyster (Crassostrea gigas), the most productive economical bivalve mollusc, is identified as an attractive model for developmental studies due to its classical mosaic developmental pattern. Myosin heavy chain is a structural and functional component of myosin, the key muscle protein of thick filament. Here, full length cDNA of striated myosin heavy chains in C. Read More

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Generation and Breeding of -Transgenic Marmoset Monkeys: Cell Chimerism and Implications for Disease Modeling.

Cells 2021 Feb 27;10(3). Epub 2021 Feb 27.

Platform Degenerative Diseases, German Primate Center-Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany.

Genetic modification of non-human primates (NHP) paves the way for realistic disease models. The common marmoset is a NHP species increasingly used in biomedical research. Despite the invention of RNA-guided nucleases, one strategy for protein overexpression in NHP is still lentiviral transduction. Read More

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February 2021

Tmem100-BAC-EGFP mice to selectively mark and purify embryonic endothelial cells of large caliber arteries in mid-gestational vascular formation.

Genesis 2021 Apr 2;59(4):e23416. Epub 2021 Mar 2.

Department of Molecular Physiology, National Cerebral and Cardiovascular Center Research Institute, Osaka, Japan.

Embryonic vascular development is achieved through the complex arrays of differentiation, proliferation, migration and mutual interaction of different cell types, and visualization as well as purification of unique cell populations are fundamental in studying its detailed mechanisms using in vivo experimental models. We previously demonstrated that Tmem100 was a novel endothelial gene encoding a small transmembrane protein, and that Tmem100 null mice showed embryonic lethality due to severe impairment of vascular formation. In the present study, we generated an EGFP reporter mouse line using a 216 kb genomic region containing mouse Tmem100 gene. Read More

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A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus.

Vet Res 2021 Feb 27;52(1):35. Epub 2021 Feb 27.

Key Laboratory of Jiangsu Preventive Veterinary Medicine, Key Laboratory for Avian Preventive Medicine, Ministry of Education, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China.

Recently, the outbreaks of hydropericardium-hepatitis syndrome (HHS) caused by the highly pathogenic fowl adenovirus serotype 4 (FAdV-4) have resulted in huge economic losses to the poultry industry globally. Although several inactivated or subunit vaccines have been developed against FAdV-4, live-attenuated vaccines for FAdV-4 are rarely reported. In this study, a recombinant virus FA4-EGFP expressing EGFP-Fiber-2 fusion protein was generated by the CRISPR/Cas9 technique. Read More

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February 2021

Valproic Acid Enhance Reprogramming of Bactrian Camel Cells through Promoting the Expression of Endogenous Gene c-Myc and the Process of Angiogenesis.

Int J Stem Cells 2021 Feb 28. Epub 2021 Feb 28.

Department of Veterinary Obstetrics, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.

Background And Objectives: Induced pluripotent stem cells (iPSCs) are usually generated by reprogramming differentiated cells throughthe introduction of specific transcription factors, but this is a difficult and inefficient process. Valproic acid (VPA) is a histone deacetylase inhibitor that significantly improves the efficiency of iPSC generation. But its role and mechanism are still unclear. Read More

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February 2021

Co-delivery of PSMA antigen epitope and mGM-CSF with a cholera toxin-like chimeric protein suppressed prostate tumor growth via activating dendritic cells and promoting CTL responses.

Vaccine 2021 Mar 19;39(11):1609-1620. Epub 2021 Feb 19.

School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, PR China. Electronic address:

Subunit vaccines derived from tumor antigens play a role in tumor therapy because of their unique advantages. However, because of the weak immunogenicity of peptides in subunit vaccines, it is difficult to trigger an effective cytotoxic T lymphocyte (CTL) response, which is critical for cancer therapy. A requirement for the activation of CTL cells by exogenous antigens is the stimulation of antigen presenting cells (APC) with the help of adjuvants and cross-presentation to T lymphocytes. Read More

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Mouse Models for Studying Hippocampal Adult Neural Stem Cell Biology.

Methods Mol Biol 2021 ;2224:61-74

Department of Pediatrics, Baylor College of Medicine, Houston, TX, USA.

The mammalian hippocampus shows a remarkable capacity for continued neurogenesis throughout life. Newborn neurons, generated by the radial neural stem cells (NSCs), are important for learning and memory as well as mood control. During aging, the number and responses of NSCs to neurogenic stimuli diminish, leading to decreased neurogenesis and age-associated cognitive decline and psychiatric disorders. Read More

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Ciliary neurotrophic factor overexpression protects the heart against pathological remodelling in angiotensin II-infused mice.

Biochem Biophys Res Commun 2021 Apr 12;547:15-22. Epub 2021 Feb 12.

Department of Cardiovascular Medicine, Key Laboratory of Heart Failure Prevention & Treatment of Hengyang, The Second Affiliated Hospital of the University of South China, Jiefang Road 30, Hengyang, 421000, Hunan, PR China. Electronic address:

Background: Ciliary neurotrophic factor (CNTF), which is a neural peptide, has been reported to confer cardioprotective effects. However, whether CNTF-based gene therapy could prevent cardiac remodelling remains incompletely clear. In this study, we used adeno-associated viral vector serotype 9 (AAV9)-based cardiac gene therapy to test the effects of CNTF overexpression on adverse ventricular remodelling in angiotensin II (Ang II)-infused mice. Read More

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Photo-dependent membrane-less organelles formed from plant phyB and PIF6 proteins in mammalian cells.

Int J Biol Macromol 2021 Apr 11;176:325-331. Epub 2021 Feb 11.

Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Sciences, 4 Tikhoretsky Ave., St. Petersburg 194064, Russian Federation; Peter the Great St.-Petersburg Polytechnic University, Polytechnicheskaya 29, St. Petersburg 195251, Russian Federation. Electronic address:

Plant photobodies are the membrane-less organelles (MLOs) that can be generated by protein-protein interactions between active form of phytochrome B (phyB) and phytochrome-interacting factors (PIFs). These organelles regulate plant photomorphogenesis. In this study, we developed two chimeric proteins with fluorescent proteins, phyB fused to EGFP and PIF6 fused to mCherry, and investigated their exogenous expression in mammalian cells by confocal fluorescence microscopy. Read More

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Experimental Infection of Calves with Transfected Attenuated Expressing the Bm86 Antigen and eGFP Marker: Preliminary Studies towards a Dual Anti-Tick/Babesia Vaccine.

Pathogens 2021 Jan 29;10(2). Epub 2021 Jan 29.

Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164-7040, USA.

Bovine babesiosis, caused by and , is a major tick-borne disease of cattle with global economic impact. The disease can be prevented using integrated control measures including attenuated vaccines, babesicidal drugs, and tick control approaches. Vaccination of cattle with the Bm86-based recombinant vaccine reduces the fitness of and , but several booster inoculations are required to maintain protection. Read More

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January 2021

Epithelial Regeneration After Doxorubicin Arises Primarily From Early Progeny of Active Intestinal Stem Cells.

Cell Mol Gastroenterol Hepatol 2021 Feb 8;12(1):119-140. Epub 2021 Feb 8.

Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina. Electronic address:

Background & Aims: aISCs (aISCs) are sensitive to acute insults including chemotherapy and irradiation. Regeneration after aISC depletion has primarily been explored in irradiation (IR). However, the cellular origin of epithelial regeneration after doxorubicin (DXR), a common chemotherapeutic, is poorly understood. Read More

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February 2021

Quantitative intravital imaging in zebrafish reveals dynamics of physiological-stress-induced mitophagy.

J Cell Sci 2021 Feb 22;134(4). Epub 2021 Feb 22.

Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA

Mitophagy, the selective recycling of mitochondria through autophagy, is a crucial metabolic process induced by cellular stress, and defects are linked to aging, sarcopenia and neurodegenerative diseases. To therapeutically target mitophagy, the fundamental dynamics and molecular mechanisms must be fully understood. Here, we generated mitophagy biosensor zebrafish lines expressing mitochondrially targeted, pH-sensitive fluorescent probes, mito-Keima and mito-EGFP-mCherry, and used quantitative intravital imaging to illuminate mitophagy during physiological stresses, namely, embryonic development, fasting and hypoxia. Read More

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February 2021

Identification of SARS-CoV-2 spike mutations that attenuate monoclonal and serum antibody neutralization.

Cell Host Microbe 2021 03 27;29(3):477-488.e4. Epub 2021 Jan 27.

Department of Molecular Microbiology, School of Medicine, Washington University in St. Louis, St. Louis, MO, USA. Electronic address:

Neutralizing antibodies against the SARS-CoV-2 spike (S) protein are a goal of COVID-19 vaccines and have received emergency use authorization as therapeutics. However, viral escape mutants could compromise efficacy. To define immune-selected mutations in the S protein, we exposed a VSV-eGFP-SARS-CoV-2-S chimeric virus, in which the VSV glycoprotein is replaced with the S protein, to 19 neutralizing monoclonal antibodies (mAbs) against the receptor-binding domain (RBD) and generated 50 different escape mutants. Read More

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Simultaneous real-time analysis of Paneth cell and intestinal stem cell response to interferon-γ by a novel stem cell niche tracking method.

Biochem Biophys Res Commun 2021 03 30;545:14-19. Epub 2021 Jan 30.

Innate Immunity Laboratory, Department of Cell Biological Science, Faculty of Advanced Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan; Innate Immunity Laboratory, Graduate School of Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan. Electronic address:

Paneth cells and Lgr5+ intestinal stem cells (Lgr5+ ISCs) constitute the stem cell niche and maintain small intestinal epithelial integrity by recognizing various niche factors derived from subepithelial cells and external antigens. Although it has been known that interferon-γ (IFN-γ), a Th1 cytokine, is associated with intestinal epithelial disruption during inflammation as a niche factor, dynamics of Paneth cells and Lgr5+ ISCs in response to IFN-γ remain to be understood. Here we show that CAG-tdTomato;Lgr5-EGFP (CT-LE) mice generated in this study enable to identify Paneth cells and Lgr5+ ISCs separately by fluorescence signals. Read More

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Cellular response to spinal cord injury in regenerative and non-regenerative stages in Xenopus laevis.

Neural Dev 2021 02 2;16(1). Epub 2021 Feb 2.

Center for Aging and Regeneration, Departamento de Biología Celular y Molecular, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Alameda 340, Santiago, Chile.

Background: The efficient regenerative abilities at larvae stages followed by a non-regenerative response after metamorphosis in froglets makes Xenopus an ideal model organism to understand the cellular responses leading to spinal cord regeneration.

Methods: We compared the cellular response to spinal cord injury between the regenerative and non-regenerative stages of Xenopus laevis. For this analysis, we used electron microscopy, immunofluorescence and histological staining of the extracellular matrix. Read More

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February 2021

A novel all-in-one conditional knockout system uncovered an essential role of DDX1 in ribosomal RNA processing.

Nucleic Acids Res 2021 04;49(7):e40

Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.

Generation of conditional knockout (cKO) and various gene-modified cells is laborious and time-consuming. Here, we established an all-in-one cKO system, which enables highly efficient generation of cKO cells and simultaneous gene modifications, including epitope tagging and reporter gene knock-in. We applied this system to mouse embryonic stem cells (ESCs) and generated RNA helicase Ddx1 cKO ESCs. Read More

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