genesis最新文献 - Book学术

Generation of Knock-In Syrian Hamsters via Zygote Microinjection Using CRISPR/Cas9 Genome Editing 使用CRISPR/Cas9基因组编辑通过受精卵显微注射产生敲入的叙利亚仓鼠

IF 2.4 4区生物学

genesis Pub Date : 2025-09-18 DOI: 10.1002/dvg.70027

Mayo Shigeta, Ken-ichi Inoue, Naoko Shimada, Alisa Tobe, Takaya Abe, Hiroshi Kiyonari

{"title":"Generation of Knock-In Syrian Hamsters via Zygote Microinjection Using CRISPR/Cas9 Genome Editing","authors":"Mayo Shigeta, Ken-ichi Inoue, Naoko Shimada, Alisa Tobe, Takaya Abe, Hiroshi Kiyonari","doi":"10.1002/dvg.70027","DOIUrl":"10.1002/dvg.70027","url":null,"abstract":"Syrian hamsters (Mesocricetus auratus) have long served as valuable model organisms in diverse research fields such as oncology, immunology, and physiology owing to their unique biological and pathological characteristics. Although embryo manipulation techniques such as embryo collection, pronuclear microinjection, and embryo transfer have been established, gene knock-in (KI) hamsters have not yet been reported. Here, we report the successful generation of gene KI Syrian hamsters by microinjecting CRISPR/Cas9 components and plasmid DNA into pronuclear-stage zygotes. Targeted insertion of a DNA cassette up to 8 kb was achieved at the ROSA26 orthologous locus and other genomic sites. Importantly, we confirmed functional expression of a reporter cassette inserted at the ROSA26 site, providing evidence of transcriptional activity at this locus in Syrian hamsters. Furthermore, we demonstrated that frozen-thawed KI embryos could give rise to live offspring using a simplified freezing and thawing protocol originally developed for mice and rats. These results confirm the feasibility and applicability of advanced genome editing technologies in Syrian hamsters. These technological advancements enable the development of versatile KI models for applications such as gene expression monitoring and conditional mutagenesis, thereby expanding the utility of Syrian hamsters as model organisms, comparable to mice and rats.","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 5","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12444848/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145082198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Generation of Astrocyte-Selective Cre Driver Lines With Distinct Onsets of Recombination Activity During Development 在发育过程中具有不同重组活性的星形细胞选择性Cre驱动系的产生

IF 2.4 4区生物学

genesis Pub Date : 2025-09-02 DOI: 10.1002/dvg.70025

Yukina Izumi, Tomoya Nakatani, Harumi Takai, Minoru Kumai, Tsutomu Kamisako, Hiroyoshi Ishizaki, Yuichi Ono

{"title":"Generation of Astrocyte-Selective Cre Driver Lines With Distinct Onsets of Recombination Activity During Development","authors":"Yukina Izumi, Tomoya Nakatani, Harumi Takai, Minoru Kumai, Tsutomu Kamisako, Hiroyoshi Ishizaki, Yuichi Ono","doi":"10.1002/dvg.70025","DOIUrl":"https://doi.org/10.1002/dvg.70025","url":null,"abstract":"<div>\u0000 \u0000 Astrocytes are a major glial cell type, playing multiple roles in the development, function, and pathogenesis of the brain. Accordingly, neuronal–astrocyte communication is an important research area. However, because these cell types share the same developmental origin, selective manipulation of each cell type is needed for precise mechanistic understanding. Here, we generated two new Cre driver lines for selective gene manipulation in astrocytes: Slc7a10-IRES-Cre and Aldh1l1-IRES-Cre. An internal ribosome entry site (IRES)-Cre cassette was knocked-in to the 3′-untranslated region of the solute carrier family 7 member 10 (Slc7a10) or aldehyde dehydrogenase 1 family member L1 (Aldh1l1) locus without disrupting gene function. The Slc7a10-IRES-Cre line underwent highly selective recombination in astrocytes of the brain, apart from choroid plexus epithelial cells. The onset of recombination began after completion of differentiation in the astrocyte lineage. By contrast, the Aldh1l1-IRES-Cre line began recombination during astrocyte differentiation at early postnatal stages. Some leaky expression was observed in the oligodendrocyte lineage, probably due to early onset of Cre expression in an uncommitted glial progenitor state. Together, the combination of the two deleter lines with distinct temporal Cre expression patterns serves as valuable tools to understand the development and function of astrocytes.\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 5","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144927675","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A Cxcl15 Cre Recombinase Mouse Model Useful to Study Gland Development in the Uterus 一种用于研究子宫腺体发育的cxcl15cre重组酶小鼠模型

IF 2.4 4区生物学

genesis Pub Date : 2025-09-01 DOI: 10.1002/dvg.70026

Andrew M. Kelleher, Hong Im Kim, Greeshma Sai Bayammagari, Daniel J. Davis, Thomas E. Spencer

{"title":"A Cxcl15 Cre Recombinase Mouse Model Useful to Study Gland Development in the Uterus","authors":"Andrew M. Kelleher, Hong Im Kim, Greeshma Sai Bayammagari, Daniel J. Davis, Thomas E. Spencer","doi":"10.1002/dvg.70026","DOIUrl":"https://doi.org/10.1002/dvg.70026","url":null,"abstract":"<div>\u0000 \u0000 The mammalian uterus contains glands in the endometrium that develop only or primarily after birth. In the mouse, endometrial glands govern post implantation pregnancy establishment via regulation of blastocyst implantation, stromal cell decidualization, and placental development. Here, we describe a new uterine glandular epithelium (GE) specific Cre recombinase mouse line that is useful to study endometrial gland development and function. Utilizing CRISPR-Cas9 genome editing, improved Cre recombinase (iCre) was inserted into the endogenous C-X-C motif chemokine ligand 15 (Cxcl15) gene. Cxcl15 mRNA, Cxcl15 protein, and Cxcl15-iCre recombinase activity were specific to the developing GE of the uterus. Cxcl15-iCre mice were crossed with floxed Foxa2 mice to conditionally delete Foxa2 specifically in the glands of the neonatal mouse uterus. This conditional deletion of Foxa2 in the developing neonatal uterus resulted in adult mice that lacked Foxa2 in the GE of the uterus, and the adult mice were infertile. The studies described here establish that Cxcl15-iCre mice are a valuable resource to elucidate and explore mechanisms regulating the development and function of glands in the uterus.\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 5","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144923519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Current Perspectives on Mesenchymal Stem Cells as a Potential Treatment for Periodontal Diseases and Conditions 间充质干细胞作为牙周病和牙周病潜在治疗方法的研究进展

IF 2.4 4区生物学

genesis Pub Date : 2025-08-21 DOI: 10.1002/dvg.70024

Shuyu Cai, Cong Li, Biaowen Wei, Ziyu Ye, Qin Mao, Wanxiang Ye, Mingdeng Rong, Jincheng Zeng

{"title":"Current Perspectives on Mesenchymal Stem Cells as a Potential Treatment for Periodontal Diseases and Conditions","authors":"Shuyu Cai, Cong Li, Biaowen Wei, Ziyu Ye, Qin Mao, Wanxiang Ye, Mingdeng Rong, Jincheng Zeng","doi":"10.1002/dvg.70024","DOIUrl":"https://doi.org/10.1002/dvg.70024","url":null,"abstract":"Periodontal diseases, including periodontitis and gingivitis, constitute a major global health burden, affecting over 1 billion people worldwide. These conditions typically initiate in adulthood and progress chronically, often exhibiting severe manifestations. The socioeconomic impact is particularly acute in low- and middle-income countries, where limited healthcare access exacerbates disease outcomes. Although conventional treatments provide symptomatic relief, they often fail to achieve complete tissue regeneration due to the complex pathophysiology of periodontal destruction. Mesenchymal stem cells (MSCs) have emerged as a transformative therapeutic strategy, demonstrating unique capabilities for immunomodulation, anti-inflammatory effects, and multipotent differentiation. Preclinical studies have documented MSC-mediated regeneration of periodontal ligaments, alveolar bone, and cementum through paracrine signaling and direct tissue integration. Clinical trials further substantiate their potential to improve key outcomes, including clinical attachment levels and probing depth reduction. However, five critical challenges require resolution for successful translation: (1) cellular source standardization, (2) mechanistic understanding of long-term efficacy, (3) safety and immunological profiling, (4) ethical and economic barriers, and (5) clinical translation barriers. This review systematically evaluates current evidence on MSC-based periodontal regeneration, analyzes these translational challenges, and provides strategic guidance for future research. By integrating fundamental science with clinical perspectives, this work advances the development of reliable MSC therapies for periodontal regeneration.","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 4","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/dvg.70024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144881055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Expression Patterns of sfrp Genes During Regeneration in the Holothurian Eupentacta fraudatrix sfrp基因在假海棠再生过程中的表达模式

IF 2.4 4区生物学

genesis Pub Date : 2025-07-29 DOI: 10.1002/dvg.70023

Konstantin Sadriev

引用次数: 0

Development of a Hepatocyte-Specific Temporal Genetic Mouse Model Using Albumin Promoter-Driven FlpER2 Expression 利用白蛋白启动子驱动的FlpER2表达建立肝细胞特异性时间遗传小鼠模型

IF 2.4 4区生物学

genesis Pub Date : 2025-07-09 DOI: 10.1002/dvg.70022

Bin Wang, Jiale Wang, Yang Liu, Gordon He, Yasmin Jahan-Mihan, Yuxi Wang, Jie Hu, Audrey Li, Faheem Muhammad, Yan Bi, Baoan Ji

{"title":"Development of a Hepatocyte-Specific Temporal Genetic Mouse Model Using Albumin Promoter-Driven FlpER2 Expression","authors":"Bin Wang, Jiale Wang, Yang Liu, Gordon He, Yasmin Jahan-Mihan, Yuxi Wang, Jie Hu, Audrey Li, Faheem Muhammad, Yan Bi, Baoan Ji","doi":"10.1002/dvg.70022","DOIUrl":"https://doi.org/10.1002/dvg.70022","url":null,"abstract":"<div>\u0000 \u0000 Tissue-specific gene manipulation using Cre/loxP or Flp/frt recombination systems is a cornerstone of genetically engineered mouse models. In this study, we aim to develop a novel hepatocyte-specific, tamoxifen-inducible Flp mouse line. BAC (bacterial artificial chromosome)-Alb(albumin)-FlpER2(estrogen receptor ligan binding domain) was developed by inserting IRES-FlpER2 cDNA between the translation stop codon and 3′-UTR of the mouse albumin gene in a bacterial artificial chromosome. Upon tamoxifen induction in mice crossed with reporter lines, western blotting, immunohistochemistry, immunofluorescence staining, and X-gal staining (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside Staining) were used to verify the recombination efficiency and specificity of this mouse model. Recombination was highly efficient and specific in hepatocytes, with no recombination detected in intrahepatic cholangiocytes or other organs in this mouse model. We generated a new tamoxifen-induced hepatocyte-specific mouse model with highly efficient recombination specifically in hepatocytes, and this model can be used to generate tumor model lines.\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 4","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-07-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144582132","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tracing Early Migratory Neurons in the Developing Nose Using Contactin-2 (Cntn2) CreERT2 利用接触蛋白-2 (Cntn2) CreERT2追踪鼻子发育中的早期迁移神经元

IF 2.4 4区生物学

genesis Pub Date : 2025-06-28 DOI: 10.1002/dvg.70021

Enrico Amato Jr., Alexis M. Semon, Paolo E. Forni

{"title":"Tracing Early Migratory Neurons in the Developing Nose Using Contactin-2 (Cntn2) CreERT2","authors":"Enrico Amato Jr., Alexis M. Semon, Paolo E. Forni","doi":"10.1002/dvg.70021","DOIUrl":"https://doi.org/10.1002/dvg.70021","url":null,"abstract":"Neuronal migration during embryonic development is a fundamental process. In the developing nose of rodents, neurons that form during early neurogenic waves in the olfactory placode leave this structure to migrate toward or into the developing brain as part of the migratory mass. This mass includes gonadotropin-releasing hormone-1 (GnRH-1) neurons, pioneer/terminal nerve (TN) neurons, as well as neural crest-derived olfactory glial cells called olfactory ensheathing cells. There have been a limited number of molecular markers available to effectively trace and functionally manipulate the early migratory neurons that originate in the olfactory region. Contactin-2 (Cntn2), also known as transiently expressed axonal surface glycoprotein-1 (TAG-1), has been used to label various developing neuronal populations, including the commissural neurons of the spinal cord, motor neurons, and TN neurons. Previous single-cell RNA sequencing analyses of the developing olfactory system have identified Cntn2 expression in the TN, suggesting that Cntn2 is a suitable molecular marker for studying nasal migratory neurons. To trace Cntn2 expression in the developing olfactory system, we generated an inducible Cntn2CreERT2 mouse line. In this study, we outline how this mouse line can serve as an effective tool for time-controlled chimeric manipulation of specific neuronal populations of interest.","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 4","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-06-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/dvg.70021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144503064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification and Classification of Zinc Finger Proteins Encoded in the Genome of an Ascidian 一种海鞘动物基因组锌指蛋白的鉴定与分类

IF 2.4 4区生物学

genesis Pub Date : 2025-06-27 DOI: 10.1002/dvg.70020

Natsuko Tamura, Yutaka Satou

引用次数: 0

Generation of Plexin-B1 Conditional Knockout Mouse With CRISPR/Cas9 Technology 用CRISPR/Cas9技术培养丛蛋白b1条件敲除小鼠

IF 2.4 4区生物学

genesis Pub Date : 2025-06-25 DOI: 10.1002/dvg.70019

Haofei Ni, Kevin Kelley, Ning Xie, Hongyan Zou, Roland H. Friedel

{"title":"Generation of Plexin-B1 Conditional Knockout Mouse With CRISPR/Cas9 Technology","authors":"Haofei Ni, Kevin Kelley, Ning Xie, Hongyan Zou, Roland H. Friedel","doi":"10.1002/dvg.70019","DOIUrl":"https://doi.org/10.1002/dvg.70019","url":null,"abstract":"<div>\u0000 \u0000 Plexins are axon guidance transmembrane receptors that control cytoskeleton and membrane dynamics in development and adult physiology. As plexins are expressed in multiple cell types in various tissues, floxed alleles that enable conditional deletion are needed to facilitate cell type-specific functional analysis. We report here the generation of a conditional floxed allele of Plexin-B1 (gene symbol Plxnb1) in mouse using CRISPR/Cas9 technology to insert two loxP sites flanking critical exons. Targeting reagents (Cas9 protein, sgRNAs, ssODNs) were delivered into single-cell embryos by electroporation. After screening a total of 128 mouse pups by PCR and Sanger sequencing, two mice were identified carrying both loxP sites in the targeted Plxnb1 locus (success rate ~ 1.6%). The usage of Alt-R modified ssODNs increased targeting frequencies at one loxP site, but not the other. We also tested homology directed repair (HDR) enhancer V2 reagent, but addition of the enhancer reduced the viability of mouse embryos. The Plxnb1flox allele was successfully transmitted through the germline in Mendelian ratios, and effective excision of the floxed region was confirmed by breeding with Cre recombinase strains.\u0000 </div>","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 3","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144482225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Genetic Mechanism That Defines the Characteristic Neurogenesis Patterns in the Neural Plate Using Hes/her Genes During Early Vertebrate Development 在脊椎动物早期发育过程中，利用he /her基因确定神经板中特征性神经发生模式的遗传机制

IF 2.4 4区生物学

genesis Pub Date : 2025-06-02 DOI: 10.1002/dvg.70015

Takero Ohyanagi, Hiroki Tokizaki, Takehisa Sato, Momo Tsuruoka, Kyo Yamasu

{"title":"Genetic Mechanism That Defines the Characteristic Neurogenesis Patterns in the Neural Plate Using Hes/her Genes During Early Vertebrate Development","authors":"Takero Ohyanagi, Hiroki Tokizaki, Takehisa Sato, Momo Tsuruoka, Kyo Yamasu","doi":"10.1002/dvg.70015","DOIUrl":"https://doi.org/10.1002/dvg.70015","url":null,"abstract":"In the early zebrafish neural plate, proneural cluster domains are defined by surrounding neural progenitor pools (NPPs), generating primary neurogenesis patterns. In each NPP, several Notch-independent Hes/her-type genes are expressed in distinct manners. Previous knockdown (KD) experiments induced ectopic neurogenesis in NPPs where only the targeted her genes were expressed, with other her genes absent, suggesting cooperative functions of Notch-independent her genes. In this study, to overcome the inherent limitations in KD approaches, we knocked out (KO) three her genes, her3, her5, and her11, using genome editing techniques. The resulting mutants exhibited ectopic neurogenesis patterns at the end of gastrulation, similar to those observed in KD experiments. KOs of her5 and her11 induced ectopic neurogenesis around the midbrain-hindbrain boundary, whereas her3 KO led to ectopic neurogenesis in rhombomere 1/2 and r4. In these cases, the expression of other Notch-independent her genes was not affected, except for her11, whose expression depended on her5. Analyses of compound mutants revealed that their phenotypes were essentially the sum of those of individual her mutants, indicating independent suppression of neurogenesis by Notch-independent her genes. In conclusion, different Notch-independent her genes collectively define the characteristic pattern of primary neurogenesis in the neural plate.","PeriodicalId":12718,"journal":{"name":"genesis","volume":"63 3","pages":""},"PeriodicalIF":2.4,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/dvg.70015","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0