A brief overview of transgenic farm animals

Transgenesis offers new possibilities to rapidly modify the genome of living organisms. The application of transgenesis to farm animals faces many problems, more than those observed in the transgenesis of laboratory animals, as there are currently many different techniques available to obtain transgenic animals, which all have problems regarding low efficiency and high costs. When these techniques are applied to farm animals the problems concerning transgenesis are multiplied. Two main techniques, male pronuclear microinjection and sperm mediated gene transfer, utilised in farm animal transgenesis, are briefly presented. The improvement of these techniques and the employment of other biotechnologies such as cloning, could expand the uses of transgenic farm animals for human health.     

Use of Transgenic Animals to Improve Human Health and Animal Production

Contents Transgenic animals are more widely used for various purposes. Applications of animal transgenesis may be divided into three major categories: (i) to obtain information on gene function and regulation as well as on human diseases, (ii) to obtain high value products (recombinant pharmaceutical proteins and xeno-organs for humans) to be used for human therapy, and (iii) to improve animal products for human consumption. All these applications are directly or not related to human health. Animal transgenesis started in 1980. Important improvement of the methods has been made and are still being achieved to reduce cost as well as killing of animals and to improve the relevance of the models. This includes gene transfer and design of reliable vectors for transgene expression. This review describes the state of the art of animal transgenesis from a technical point of view. It also reports some of the applications in the medical field based on the use of transgenic animal models. The advance in the generation of pigs to be used as the source of organs for patients and in the preparation of pharmaceutical proteins from milk and other possible biological fluids from transgenic animals is described. The projects in course aiming at improving animal production by transgenesis are also depicted. Some the specific biosafety and bioethical problems raised by the different applications of transgenesis, including consumption of transgenic animal products are discussed.     

转基因在动物生产中的应用

人们一直期望畜产品和人类健康产品能快速得到提高,DNA重组技术和转基因的出现使得在不同物种间,甚至不同系统发育领域间的这一提高在很大范围内变为可能。如今,我们能在细菌中生产人类胰岛素,在牛奶中获得人类凝结因子。转基因、动物克隆和动物多产技术的进步在一定程度上已经实现了在动物转基因领域的期望。作者回顾了当前动物转基因乳、肉和转基因抗病动物的研究近况,并讨论了一些由转基因技术应用而引发的生物伦理学和商业性问题。     

Transgenesis may affect farm animal welfare: a case for systematic risk assessment

This paper considers (potentially) harmful consequences of transgenesis for farm animal welfare and examines the strategy of studying health and welfare of transgenic farm animals. Evidence is discussed showing that treatments imposed in the context of farm animal transgenesis are by no means biologically neutral and may compromise animal health and welfare. Factors posing a risk for the welfare of transgenic farm animals include integration of a transgene within an endogenous gene with possible loss of host gene function (insertional mutations), inappropriate transgene expression and exposure of the host to biologically active transgene-derived proteins, and in vitro reproductive technologies employed in the process of generating transgenic farm animals that may result in an increased incidence of difficult parturition and fetal and neonatal losses and the development of unusually large or otherwise abnormal offspring (large offspring syndrome). Critical components of a scheme for evaluating welfare of transgenic farm animals are identified, related to specific characteristics of transgenic animals and to factors that may interact with the effects of transgenesis. The feasibility of an evaluation of welfare of transgenic farm animals in practice is addressed against the background of the objectives and conditions of three successive stages in a long-term transgenic program. Concrete steps with regard to breeding and testing of transgenic farm animals are presented, considering three technologies to generate transgenic founders: microinjection, electroporation and nuclear transfer, and gene targeting including gene knockout. The proposed steps allow for unbiased estimations of the essential treatment effects, including hemi- and homozygous transgene effects as well as effects of in vitro reproductive technologies. It is suggested that the implementation of appropriate breeding and testing procedures should be accompanied by the use of a comprehensive welfare protocol, specifying which parameters to monitor, at which stages of the life of a farm animal, and in how many animals. Some prerequisites and ideas for such a protocol are given. It is anticipated that systematic research into the welfare of farm animals involved in transgenesis will facilitate the use of the safest experimental protocols as well as the selection and propagation of the healthiest animals and, thereby, enable technological progress that could be ethically justified.     

mRNA技术及其在动物疫病研究中的应用进展

mRNA技术是一种将人工合成的mRNA分子引入细胞内，并利用细胞自身的翻译机制将其翻译成蛋白质的技术。作为一种新兴的生物技术，mRNA技术已经在疫苗研究、基因治疗等多个领域取得了重要进展，并在新冠预防和传染病防治方面进行了临床应用，展现出了广阔的应用前景。通过对mRNA技术的发展历程，mRNA疫苗的设计、递送及特点，以及其在动物疫病研究的进展予以综述，分析其存在的问题，展望今后的发展方向，为相关研究提供参考。     

New Biotechniques and their Consequences for Farm Animal Welfare

Contents This paper considers (potentially) harmful consequences of new biotechnologies for farm animal welfare. The most important new biotechnologies that are currently used in farm animals breeding and husbandry include: multiple ovulation and embryo transfer (MOET) and in vitro embryo production (IVP). Cloning by nuclear transfer (NT) and transgenesis are still in development and mainly applied for experimental purposes with the prospect of a more widespread practical implemention in the future. Evidence is presented showing that generally accepted technologies such as MOET and IVP, relative to in vivo procedures, can result in a host of deleterious side-effects commonly known as the large offspring syndrome (LOS). Likewise, NT and transgenesis, which also typically include several in vitro reproductive manipulations, have clearly been associated with the occurrence of LOS symptoms. It is argued that transgenesis may constitute one additional set of factors that may negatively affect farm animal welfare: the expression of the transgene and the concomitant synthesis and release of a protein. NT might lead to incompletely reprogramming of the transferred genome. It is suggested that the introduction of new biotechnologies into farm animal husbandry should be accompanied by scientifically valid and systematic studies into the effects on animal welfare, with the help of a comprehensive welfare protocol.     

昆虫转座子及在家蚕中的应用

概述了近年来广泛应用于昆虫的若干转座子及其基本构造和应用;着重讨论了piggyBac转座子的构造和转座机制及所作成的转基因昆虫种类及标记基因:同时列举了在家蚕转基因中的实例.     

基因治疗中的非病毒型载体研究进展

基因治疗载体一般分为病毒性载体和非病毒性载体。由于病毒型载体存在安全性问题,目前非病毒性载体的研究越来越受到人们的重视。非病毒载体是新兴的基因转导系统,具有低毒、低免疫反应、外源基因整合几率低、无基因插入片断大小限制,以及使用简单、制备方便、便于保存和检验等优势。脂质体作为常用的非病毒性载体容易制备,安全性高,却难达到质控要求,体内基因导入效率低,且无靶向性,使得其应用受到限制。pH一敏脂质体、阳离子脂质体等大大提高了脂质体的转运效率。同时高分子载体、纳米基因转运体在基因治疗中的应用,也为非病毒载体在临床上的应用开辟了广阔的前景。     

转基因山羊研究进展

近年来,动物转基因技术发展日淅完善,在牛、羊、猪等家畜上的应用日益增多.为全面了解山羊转基因技术的研究进展,论文简要回顾了国内外转基因山羊生产的里程碑事件,总结了生产转基因山羊的方法和原理,并对转基因山羊研究状况、应用前景及存在问题进行了较为系统的概括和分析,从而为借助现代生物技术丰富我国山羊育种手段,加快山羊新良种培...     

精子载体转基因的研究进展

精子载体法培养转基因动物是目前操作最简便，成本低廉且危险隐患相对较低的一种转基因方法。近10年来的研究结果表明，精子载体法可得到转基因阳性动物，并且引起了更多科研人员的关注。     

影响猪精子介导基因转移效果的因素研究

为优化猪精子载体法技术参数,利用荧光定量PCR、荧光显微镜检测和精液常规检测方法,分析不同DNA转染剂、不同孵育温度和不同形态DNA对猪精子转染外源DNA的影响。结果表明,PEI和TransFast转染剂能极显著提高猪精子转染效果,且PEI优于TransFast,而NanoFect转染剂包裹DNA不能转染精子。随着转染温度的升高,精子的活力和活率均明显下降,而转染率和内化外源DNA量基本保持稳定,而吸附外源DNA量呈下降趋势。环状DNA和线性化DNA在与精子共孵育后,两者的精子活力、活率、转染率和内化外源DNA量差异均不显著,精子吸附线性化DNA量极显著高于环状DNA量。综合分析表明,外源DNA形态对猪精子转染效果无显著影响;PEI和TransFast能够显著提高猪精子转染效果;共孵育温度以17℃为最佳,本结果为开展精子载体法制备转基因猪研究提供了基础试验依据。     

基因敲除和RNAi在畜牧兽医领域的研究及展望

进入21世纪,生命科学已经进入后基因组时代,大规模的基因功能研究正成为生命科学研究的热点,基因组技术的重要性也越来越明显。论述了基因敲除和RNA干涉两方面技术,并对其在畜牧兽医领域的应用进行了展望,提出了利用基因敲除和RNA干涉技术,能够更清楚地研究和发现基因新的功能,同时还可以用于制备疾病动物模型,用于科学研究和新药的筛选,为畜牧兽医带来广大的前景。     

SMAD3基因在畜牧生产中的应用研究进展

SMAD3基因是SMADS基因家族成员之一,其编码的SMAD3蛋白是转化生长因子-β(TGF-β)超家族特异的细胞内信号转导分子,SMAD3基因通过TGF-β超家族参与疾病、免疫调节、生长发育、创伤愈合、软骨与骨骼的发育及维持等重要的生理过程,同时还参与调节生殖细胞的增殖、分化、成熟、黏附、闭锁、凋亡和甾体激素的产生等,因此,了解SMAD3基因的结构与功能对动物生长发育、繁殖等研究具有重要意义。文章简述了SMAD3基因结构、功能、作用机理,分析了其在畜牧生产方面相关的应用研究进展发现,目前SMAD3基因在畜牧生产上的应用研究主要集中在参与调控脊椎动物(猪、牛、羊、鸡等)生长、发育、细胞免疫与凋亡、激素分泌及繁殖功能方面;SMAD3基因在调控动物机体疾病发生、生长发育、脂肪沉积及繁殖性能方面仍是国内外研究的热点,而该基因对畜禽疾病发生、生长和繁殖性能的精确分子调控机制仍有待进一步研究。     

Germ cells and transgenesis in chickens

Chickens have proven to be useful organisms for transgenic research. This work provides enormous benefits in advancing animal biotechnology and aids in the development of unique technologies for bioreactor production and experimental model development. The various advantages of chicken transgenesis are derived from the genetic and physiological characteristics of this organism, although several physiological properties have impeded the development of an efficient transgenic system. We have developed embryo-mediated and testis-mediated transgenic systems using chicken primordial germ cells (PGCs) from embryos and testicular cells from adult males. These methods are efficient and involve minimal technical effort. Here, we review previous transgenic research using PGCs and testicular cells from chickens. Furthermore, we have summarized the development of the chicken model system in biomedical science and biotechnology and our recent achievements in this field.     

Assisted reproductive techniques for cattle breeding in developing countries: a critical appraisal of their value and limitations

Commercialization of animal biotechnologies, including those related to reproduction [also known as assisted reproductive techniques (ARTS)], is an increasing reality in developing countries, following the enormous flow of information around us and the increasing global commercial interests in areas where cattle production has its major assets. The present review discusses the achievements of various biotechnological tools for reproduction in cattle including semen handling for artificial insemination (AI), superovulation and embryo transfer (MOET), in vitro handling of oocytes and production of embryos, reproductive cloning and emerging technologies (sex selection, gene targeting and nuclear transfer for livestock transgenesis, genomics for marker-assisted selection, etc.). The application of these technologies for cattle breeding is critically discussed in relation to their impact in the improvement of the efficiency of dairy and beef production in developed and - particularly - in developing countries, which ultimately rule the possibilities of a competitive and sound production of food for human consumption. Despite the remarkable progress made and the punctual importance of some of the above-mentioned technologies, AI remains the most important assisted reproductive technology (ART) in developing countries. Any attempt to gain widespread of any other ART under the predominant economical conditions in developing countries ought to match the simplicity and the success of AI as a breeding tool.     

角蛋白家族基因及其与动物毛发性状关系的研究进展

角蛋白基因家族由众多结构和功能相似的基因组成,分为角蛋白中间丝蛋白(KIF)和角蛋白关联蛋白(KAP)2个基因家族。角蛋白是动物毛发的结构蛋白,决定毛发纤维的基本特征,是在寻找控制毛皮性状主效基因时的首选候选基因。角蛋白基因的表达与遗传多样性有关,角蛋白的多态性与毛皮动物的毛发性状具有相关性。本文从角蛋白基因的结构与定位、多态性与毛发性状、表达差异与毛发性状、基因表达调控等方面对毛皮动物角蛋白基因的研究进展进行综述,为进一步研究角蛋白在动物生产中与经济性状的关系提供理论基础。     

Wip1基因对动物繁殖及免疫的调节作用

野生型p53诱导的磷酸酶1（wild-type p53-induced phosphatase 1,Wip1）是蛋白磷酸酶2C （protein phosphatase type 2C,PP2C）家族中的一员,可以靶向调控机体内多种重要的信号分子,如p53、MAPK和Chk1/Chk2等,在动物细胞周期、增殖、分化、凋亡、衰老、自噬及DNA损伤修复等生理过程中发挥重要作用。Wip1基因缺失会使小鼠生殖激素水平失衡,且该基因通过ATM、Wnt、凋亡和炎症等信号通路影响精子生成过程,导致雄性动物繁殖力下降。此外,Wip1基因还可通过动态平衡调节DNA损伤反应和去磷酸化作用来影响卵母细胞和胚胎发育,从而调控雌性动物的生殖。免疫系统是机体执行免疫应答及免疫功能的重要系统,其与炎症反应和肿瘤发生有着紧密的联系。Wip1基因缺失会使病原体敏感性增强,影响T细胞、B细胞和中性粒细胞迁移及凋亡,进而导致炎症反应。作为原癌基因,Wip1基因通过调控各种信号分子影响DNA损伤修复、细胞周期进程及细胞凋亡等,参与肿瘤发生。因此,Wip1基因在动物繁殖调控和免疫调节中扮演着重要角色。目前,Wip1基因受到越来越多学者的关注,特别是其调控动物疾病发生发展的机制已成为研究热点。本研究主要综述了Wip1基因对动物繁殖及免疫的调节作用,以期为家畜育种、疾病防治及靶向治疗提供新思路。     

Recombinant DNA, gene transfer and the future of animal agriculture

Recombinant deoxyribonucleic acid (DNA) technology and gene transfer are two areas of biotechnology that will have significant impact on animal agriculture. Applications to animal agriculture can be expected in animal health management, improved crops and feeds, manipulation of animal physiology, and genetic improvement of livestock species. Improved diagnostic reagents and vaccines that will improve herd health are currently under development. Yield of crop plants such as corn will be increased and the nutritional value of these feeds improved through applications of recombinant DNA technology. Administration of exogenous hormones synthesized by bacteria holds great promise for increasing the yield of milk and possibly meat. Research on the transfer of cloned genes into animals has progressed rapidly and has recently been accomplished in sheep and swine. Tissue-specific and developmentally regulated expression of transferred genes now seems possible with defined gene promoter sequences. Several applications of these biotechnologies can be expected within 5 to 10 yr, whereas others may require longer periods of research. The 21st century will herald a new era in animal science research and applications, with recombinant DNA and gene transfer playing major roles.     

脂肪酸结合蛋白（FABPs）基因多态性对畜产品品质的影响

脂肪酸结合蛋白（FABPs）属于脂结合蛋白超家族成员，广泛存在于动物细胞质中，承担着细胞内脂肪酸的运输任务。研究发现，脂肪酸结合蛋白（FABPs）基因多态性可以通过调控脂肪代谢来改善畜产品品质，提高饲料利用率，降低环境污染。就FABPs的结构和类型，FABPs对脂肪代谢的影响，FABPs基因多态性及其应用后在改善畜产品品质中发挥的作用、功效和应用特点进行了阐述，以期为改进畜禽养殖技术提供参考。     

乳蛋白基因在转基因动物乳腺细胞中特异性表达的研究进展

乳蛋白是乳的主要营养成分，乳蛋白的种类及其在乳中的浓度都影响乳的品质，而乳中各乳蛋白的含量都受到相应乳蛋白基因的控制。通过转基因技术，可以在转基因动物乳腺细胞中特异性地表达目的蛋白，从而改善乳的品质以及生产具有特殊药用价值的乳产品。本文主要概述乳蛋白多态性及其作用，乳蛋白基因及多态性，并着重介绍乳蛋白基因在转基因动物体内的表达调控；通过介绍大量转基因试验的研究成果，以探寻改变乳成分的分子遗传基础。