Rocha-Martins Maurício, Cavalheiro Gabriel R, Matos-Rodrigues Gabriel E, Martins Rodrigo A P
Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, BR.
An Acad Bras Cienc. 2015 Aug;87(2 Suppl):1323-48. doi: 10.1590/0001-3765201520140710.
Genome modification technologies are powerful tools for molecular biology and related areas. Advances in animal transgenesis and genome editing technologies during the past three decades allowed systematic interrogation of gene function that can help model how the genome influences cellular physiology. Genetic engineering via homologous recombination (HR) has been the standard method to modify genomic sequences. Nevertheless, nuclease-guided genome editing methods that were developed recently, such as ZFN, TALEN and CRISPR/Cas, opened new perspectives for biomedical research. Here, we present a brief historical perspective of genome modification methods, focusing on transgenic mice models. Moreover, we describe how new techniques were discovered and improved, present the paradigm shifts and discuss their limitations and applications for biomedical research as well as possible future directions.
基因组修饰技术是分子生物学及相关领域的强大工具。在过去三十年中，动物转基因和基因组编辑技术的进展使得对基因功能进行系统性研究成为可能，这有助于模拟基因组如何影响细胞生理学。通过同源重组（HR）进行基因工程一直是修饰基因组序列的标准方法。然而，最近开发的核酸酶引导的基因组编辑方法，如锌指核酸酶（ZFN）、转录激活因子样效应物核酸酶（TALEN）和规律成簇间隔短回文重复序列/CRISPR相关蛋白（CRISPR/Cas），为生物医学研究开辟了新的前景。在此，我们简要介绍基因组修饰方法的历史概况，重点关注转基因小鼠模型。此外，我们描述了新技术是如何被发现和改进的，呈现了范式转变，并讨论了它们在生物医学研究中的局限性和应用以及可能的未来发展方向。
