Ratner Laura Daniela, La Motta Gaston Emilio, Briski Olinda, Salamone Daniel Felipe, Fernandez-Martin Rafael
Laboratorio Biotecnología Animal (LabBA), Departamento de Producción Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, Argentina.
Instituto de Investigaciones en Producción Animal (INPA), CONICET-Universidad de Buenos Aires, Buenos Aires, Argentina.
Front Genet. 2021 Mar 5;11:617850. doi: 10.3389/fgene.2020.617850. eCollection 2020.
Pigs are an important resource for meat production and serve as a model for human diseases. Due to their physiological and anatomical similarities to humans, these animals can recapitulate symptoms of human diseases, becoming an effective model for biomedical research. Although, in the past pig have not been widely used partially because of the difficulty in genetic modification; nowadays, with the new revolutionary technology of programmable nucleases, and fundamentally of the CRISPR-Cas9 systems, it is possible for the first time to precisely modify the porcine genome as never before. To this purpose, it is necessary to introduce the system into early stage zygotes or to edit cells followed by somatic cell nuclear transfer. In this review, several strategies for pig knock-out gene editing, using the CRISPR-Cas9 system, will be summarized, as well as genotyping methods and different delivery techniques to introduce these tools into the embryos. Finally, the best approaches to produce homogeneous, biallelic edited animals will be discussed.
猪是肉类生产的重要资源，也是人类疾病的模型。由于它们在生理和解剖学上与人类相似，这些动物能够重现人类疾病的症状，成为生物医学研究的有效模型。尽管过去猪没有被广泛使用，部分原因是基因改造困难；但如今，随着可编程核酸酶这一革命性新技术，尤其是CRISPR-Cas9系统的出现，首次有可能以前所未有的精度对猪基因组进行精确改造。为此，有必要将该系统导入早期受精卵，或对细胞进行编辑后再进行体细胞核移植。在这篇综述中，将总结使用CRISPR-Cas9系统进行猪基因敲除编辑的几种策略，以及基因分型方法和将这些工具导入胚胎的不同递送技术。最后，将讨论生产同质双等位基因编辑动物的最佳方法。
