Transgene

转基因
  • 文章类型: Journal Article
    小胶质细胞在许多神经病理学的病因学中起着核心作用。转基因工具是获得对小胶质细胞功能的可靠和特异性控制的强大实验方法。腺相关病毒(AAV)载体已经是神经科学研究中不可或缺的工具。尽管广泛使用AAV,并且对小胶质细胞在中枢神经系统(CNS)功能和疾病研究中的作用有很大的兴趣,使用AAV的小胶质细胞转导很少报道。这篇综述探讨了使用AAV在小胶质细胞中表达转基因的挑战和进展。首先,我们将检查AAV衣壳的功能解剖,这将作为后续研究探索衣壳突变与小胶质细胞转导功效之间关系的讨论的基础。在概述了AAV的功能解剖结构之后,我们将考虑证明AAV介导的小胶质细胞和小胶质细胞样细胞系转导的实验证据,然后检查文献中确定的最有希望的实验方法。最后,在AAV实验方法的未来应用中,将考虑技术局限性。
    Microglia play a central role in the etiology of many neuropathologies. Transgenic tools are a powerful experiment approach to gain reliable and specific control over microglia function. Adeno-associated virus (AAVs) vectors are already an indispensable tool in neuroscience research. Despite ubiquitous use of AAVs and substantial interest in the role of microglia in the study of central nervous system (CNS) function and disease, transduction of microglia using AAVs is seldom reported. This review explores the challenges and advancements made in using AAVs for expressing transgenes in microglia. First, we will examine the functional anatomy of the AAV capsid, which will serve as a basis for subsequent discussions of studies exploring the relationship between capsid mutations and microglia transduction efficacy. After outlining the functional anatomy of AAVs, we will consider the experimental evidence demonstrating AAV-mediated transduction of microglia and microglia-like cell lines followed by an examination of the most promising experimental approaches identified in the literature. Finally, technical limitations will be considered in future applications of AAV experimental approaches.
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  • 文章类型: Journal Article
    通过基因操作开发的转基因蚊子,为可持续和有效控制蚊媒疾病提供了一个有前途的战略。这种策略依赖于大量释放转基因蚊子到野外,它们的转基因有望在自然环境中持续存在,永久或暂时,在蚊子种群中。在这种情况下,转基因蚊子的适应度是决定它们在野外生存的重要因素。转基因表达的影响,插入诱变,与实验室适应有关的近亲繁殖抑郁症,与发展纯合蚊子系有关的搭便车效应都可以对转基因蚊子的适应性产生影响。因此,转基因相关的健身成本的实时估计是必要的建模和规划转基因蚊子释放计划。这可以通过直接比较转基因的纯合或半合子个体及其野生型对应物的适应性参数来实现。或通过笼式入侵实验来监测多代转基因等位基因的频率。最近的进步,如特定地点的集成系统和基因驱动,提供平台来解决转基因蚊子的健康问题。需要对转基因个体的适应性进行更多的研究,以开发具有低适应性成本的转基因蚊子。
    Transgenic mosquitoes developed by genetic manipulation, offer a promising strategy for the sustainable and effective control of mosquito-borne diseases. This strategy relies on the mass release of transgenic mosquitoes into the wild, where their transgene is expected to persist in the natural environment, either permanently or transiently, within the mosquito population. In such circumstances, the fitness of transgenic mosquitoes is an important factor in determining their survival in the wild. The impact of transgene expression, insertional mutagenesis, inbreeding depression related to laboratory adaptation, and the hitchhiking effect involved in developing homozygous mosquito lines can all have an effect on the fitness of transgenic mosquitoes. Therefore, real-time estimation of transgene-associated fitness cost is imperative for modeling and planning transgenic mosquito release programs. This can be achieved by directly comparing fitness parameters in individuals homozygous or hemizygous for the transgene and their wild-type counterparts, or by cage invasion experiments to monitor the frequency of the transgenic allele over multiple generations. Recent advancements such as site-specific integration systems and gene drives, provide platforms to address fitness issues in transgenic mosquitoes. More research on the fitness of transgenic individuals is required to develop transgenic mosquitoes with a low fitness cost.
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  • 文章类型: Journal Article
    在过去的几十年里,基因治疗作为癌症等疾病的一种有希望的治疗策略,在医学研究中获得了巨大的重要性,艾滋病,老年痴呆症,和许多遗传性疾病。当一个基因需要被传递到人体内的靶细胞时,它必须通过细胞外和细胞内环境的大量屏障。这就是为什么裸基因和核酸的传递是非常不利的,基因传递需要合适的载体,可以将基因货物携带到靶位点并保护其免受生物降解。迄今为止,医学研究提出了两种类型的基因传递载体,它们是病毒和非病毒载体。病毒保护转基因免受生物降解的能力及其有效跨越细胞屏障的能力已允许基因治疗研究开发利用病毒及其不同基因组作为基因递送载体的新方法。尽管病毒载体非常有效,科学还提出了许多基于阳离子脂质的非病毒系统,阳离子聚合物,和无机颗粒,提供可持续的基因表达,而不会引发不必要的炎症和免疫反应,被认为是无毒的。在这次审查中,我们将详细讨论用于基因传递的所有病毒和非病毒载体的最新数据.介绍了基于病毒和非病毒载体的基因传递机制,并给出了各类载体的优缺点。
    Over the past few decades, gene therapy has gained immense importance in medical research as a promising treatment strategy for diseases such as cancer, AIDS, Alzheimer\'s disease, and many genetic disorders. When a gene needs to be delivered to a target cell inside the human body, it has to pass a large number of barriers through the extracellular and intracellular environment. This is why the delivery of naked genes and nucleic acids is highly unfavorable, and gene delivery requires suitable vectors that can carry the gene cargo to the target site and protect it from biological degradation. To date, medical research has come up with two types of gene delivery vectors, which are viral and nonviral vectors. The ability of viruses to protect transgenes from biological degradation and their capability to efficiently cross cellular barriers have allowed gene therapy research to develop new approaches utilizing viruses and their different genomes as vectors for gene delivery. Although viral vectors are very efficient, science has also come up with numerous nonviral systems based on cationic lipids, cationic polymers, and inorganic particles that provide sustainable gene expression without triggering unwanted inflammatory and immune reactions, and that are considered nontoxic. In this review, we discuss in detail the latest data available on all viral and nonviral vectors used in gene delivery. The mechanisms of viral and nonviral vector-based gene delivery are presented, and the advantages and disadvantages of all types of vectors are also given.
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  • 文章类型: Journal Article
    Genome editing is a genetic engineering technique that uses site-directed cleavage activity of specific artificial nucleases and endogenous DNA damage repair activity to generate insertions, deletions or substitutions in the targeted genomic loci. As the accuracy and efficiency of genome editing is improving and the operation is simple, the application of genome editing is expanding. This article provides an overview of the three major genome editing technologies and genome editing types, and the regulatory frameworks for genome-edited products were summarized in the United States, the European Union, and other countries. At the same time, based on the Chinese safety management principles and systems for genetically modified organisms (GMOs), the authors proposed a regulatory framework for genome-edited products. Genome-edited products should first be classified according to whether containing exogenous genetic components such as Cas9 editing enzymes or not. They should be regulated as traditional genetically modified organisms if they do. Otherwise, the regulation of genome-edited products depends on targeted modifications.
    基因组编辑技术是指利用特异性核酸酶的定点剪切活性与细胞内源DNA 损伤修复活性,在基因组水平上对目的核酸序列或单个核苷酸进行定点修饰的基因工程技术,该技术可以对生物体基因组进行精确敲除、插入、单碱基突变或置换等编辑。目前,基因组编辑技术具有精确性、高效性及易操作性,应用范围日益扩大。文中简要概述了3 种主要的基因组编辑技术工具及基因组编辑类型,介绍了美国、欧盟等国家和地区对基因组编辑产品的监管体制。同时,基于我国对转基因产品的安全管理原则与体系,初步提出了基因组编辑产品的安全管理思路。根据中间材料或产品中是否含有外源编辑酶蛋白基因成分对基因组编辑产品进行分类管理,含有外源编辑酶的材料应按现有转基因安全管理办法进行管理;中间材料或产品中不含有Cas9 等编辑酶的材料应根据被编辑位点的特征进行具体分类管理。.
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