A novel technique for large-fragment knock-in animal production without ex vivo handling of zygotes

Scientific Reports, Год журнала: 2023, Номер 13(1)

Опубликована: Фев. 8, 2023

Abstract CRISPR/Cas-based genome editing has dramatically improved genetic modification technology. In situ electroporation called via oviductal nucleic acid delivery (GONAD), which eliminates the need for ex vivo embryo handling, is technically simplest method gene transfer and can be performed in laboratories without developmental engineering expertise including micromanipulation techniques. However, use of this remains challenging case large-fragment knock-in, such as expression cassettes. Adeno-associated viruses (AAV) act donor DNA homologous recombination infected cells, rodent embryos. study, we demonstrated simultaneous AAV donors CRISPR/Cas9 components into embryos to create knock-in animals, successfully generated rats carrying a cassette with length 3.0 kb using small number animals electroporation. These findings indicate that technique an efficient high-throughput strategy producing genetically modified rodents may applicable other animal species.

Язык: Английский

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Silc1 long noncoding RNA is an immediate-early gene promoting efficient memory formation

Cell Reports, Год журнала: 2023, Номер 42(10), С. 113168 - 113168

Опубликована: Сен. 23, 2023

Long noncoding RNAs (lncRNAs) are expressed in many brain circuits and types of neurons; nevertheless, their functional significance for normal functions remains elusive. Here, we study the central nervous system Silc1, an lncRNA have shown previously to be important neuronal regeneration peripheral system. We found that Silc1 is rapidly strongly induced hippocampus upon exposure novelty required efficient spatial learning. production induction Sox11 (its cis-regulated target gene) throughout CA1-CA3 regions proper expression key genes. Consistent with its role plasticity, levels decline during aging models Alzheimer's disease. Overall, describe a plasticity pathway which acts as immediate-early gene activate induce growth-associated transcriptional program

Язык: Английский

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Genetically modified mice as a tool for the study of human diseases

Molecular Biology Reports, Год журнала: 2024, Номер 51(1)

Опубликована: Янв. 18, 2024

Язык: Английский

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Endothelial Cell-Selective Adhesion Molecule Contributes to the Development of Definitive Hematopoiesis in the Fetal Liver

Stem Cell Reports, Год журнала: 2019, Номер 13(6), С. 992 - 1005

Опубликована: Дек. 1, 2019

Endothelial cell-selective adhesion molecule (ESAM) is a lifelong marker of hematopoietic stem cells (HSCs). Although we previously elucidated the functional importance ESAM in HSCs stress-induced hematopoiesis adults, it unclear how affects development during fetal life. To address this issue, analyzed fetuses from conventional or conditional ESAM-knockout mice. Approximately half ESAM-null died after mid-gestation due to anemia. RNA sequencing analyses revealed downregulation adult-type globins and Alas2, heme biosynthesis enzyme, livers. These abnormalities were attributed malfunction HSCs, which was demonstrated culture transplantation experiments. crosslinking directly influenced gene transcription observations critical involvement expressed endothelial lethality. Thus, showed that had important roles developing definitive hematopoiesis. Furthermore, unveiled process.

Язык: Английский

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Utilization of ethanolamine phosphate phospholyase as a unique astrocytic marker

Frontiers in Cellular Neuroscience, Год журнала: 2023, Номер 17

Опубликована: Янв. 30, 2023

Astrocytes play diverse roles in the central nervous system (CNS) both physiological and pathological conditions. Previous studies have identified many markers of astrocytes to analyze their complicated roles. Recently, closure critical period by mature has been revealed, need for finding astrocyte-specific growing. We previously found that Ethanolamine phosphate phospholyase (Etnppl) was almost not expressed developing neonatal spinal cord, its expression level slightly decreased after pyramidotomy adult mice, which showed weak axonal sprouting, suggesting negatively correlates with elongation. Although Etnppl is known, utility as an astrocytic marker yet investigated detail. Here, we selectively adult. Re-analyses using published RNA-sequencing datasets revealed changes cord injury, stroke, or systemic inflammation models. produced high-quality monoclonal antibodies against ETNPPL characterized localization mice. Expression very except ventricular subventricular zones, it heterogeneously highest cerebellum, olfactory bulb, hypothalamus lowest white matter. Subcellular dominant nuclei cytosol minor population. Using antibody, were labeled cerebral cortex detected pyramidotomy. a subset Gjb6+ cord. The created, well fundamental knowledge this study, will be valuable resources scientific community expand our understanding responses conditions future analyses.

Язык: Английский

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