Houdebine Louis-Marie
Biologie du Développement et Biotechnologies, Institut National de la Recherche Agronomique, 78352 Jouy en JosasCedex, France.
J Biotechnol. 2002 Sep 25;98(2-3):145-60. doi: 10.1016/s0168-1656(02)00129-3.
Transgenic animals have been used for years to study gene function and to create models for the study of human diseases. This approach has become still more justified after the complete sequencing of several genomes. Transgenic animals are ready to become industrial bioreactors for the preparation of pharmaceuticals in milk and probably in the future in egg white. Improvement of animal production by transgenesis is still in infancy. Despite its intensive use, animal transgenesis is still suffering from technical limitations. The generation of transgenics has recently become easier or possible for different species thanks to the use of transposons or retrovirus, to incubation of sperm which DNA followed by fertilization by intracellular sperm injection or not and to the use of the cloning technique using somatic cells in which genes have been added or inactivated. The Cre-LoxP system is more and more used to withdraw a given sequence from the genome or to target the integration of a foreign DNA. The tetracycline system has been improved and can more and more frequently be used to obtain faithful expression of transgenes. Several tools: RNA forming a triple helix with DNA, antisense RNA including double strand RNA inducing RNA interference and ribozymes, and also expression of proteins having a negative transdominant effect, are tentatively being improved to inhibit specifically the expression of host or viral genes.All these techniques are expected to offer experimenters new and more precise models to study gene function even in large animals. Improvement of breeding by transgenesis has become more plausible including through the precise allele replacement in farm animals.
多年来,转基因动物一直被用于研究基因功能,并创建人类疾病研究模型。在多个基因组完成测序之后,这种方法变得更加合理。转基因动物即将成为用于在乳汁中制备药物的工业生物反应器,未来可能还会用于在蛋清中制备药物。通过转基因技术提高动物生产水平仍处于起步阶段。尽管转基因技术被广泛应用,但动物转基因技术仍存在技术局限性。由于使用转座子或逆转录病毒、对携带DNA的精子进行孵育然后通过或不通过胞内精子注射进行受精,以及使用添加或灭活了基因的体细胞克隆技术,最近不同物种转基因动物的产生变得更加容易或成为可能。Cre-LoxP系统越来越多地用于从基因组中去除特定序列或靶向外源DNA的整合。四环素系统已经得到改进,越来越频繁地用于实现转基因的忠实表达。几种工具:与DNA形成三链螺旋的RNA、包括诱导RNA干扰的双链RNA和核酶的反义RNA,以及具有负显性效应的蛋白质的表达,正在尝试改进以特异性抑制宿主或病毒基因的表达。所有这些技术有望为实验者提供新的、更精确的模型,以便即使在大型动物中也能研究基因功能。通过转基因技术改良育种变得更加可行,包括在农场动物中进行精确的等位基因替换。