靶向基因修饰技术在免疫缺陷动物模型研究中的应用进展

摘要

免疫缺陷动物模型在人类相关疾病机理研究、药物研发、器官移植研究和干细胞研究中有重要的应用。但由于传统基因修饰动物构建技术难度大、效率低等限制，在中型、大型动物中获得的免疫缺陷动物模型还很少。近年来新兴的靶向基因修饰技术，包括ZFNs、TALENs、CRISPR／Cas9等，为高效率免疫缺陷动物模型构建提供了技术基础。本文就ZFNs、TALENs、CRISPR／Cas9的技术原理及研究进展进行概况介绍，并详细地阐述这些技术在中型和大型动物中免疫缺陷动物模型构建方面取得的进展，包括KO Rag1／Rag2兔、KO Rag1／Rag2猪、KO IL2rg猪、KO Ppar-g／Rag1猴等。这些免疫缺陷动物模型不仅能用于人类SCID相关疾病研究，评价干细胞移植的效率和安全性，且可进行临床前手术治疗研究，生产人源化动物模型等，进而架起实验动物与医学应用的桥梁，促进临床前细胞再生策略的综合评价体系的发展。%The established immunodeficient animal models could be used as valuable resource for mechanism re-search of related disease in humans, drug discovery and development, translational research and stem cell research.How-ever, it is difficult and low-efficient to establish the genetic modified animal models using traditional technologies.The re-ports for immunodeficient animal models are few in middle-size and large animals.Recently, several effective gene-targeting tools, including ZFNs, TALENs, CRISPR/Cas9, develop quickly and provide technology basis for the establishment of im-munodeficient animal models.In this paper, the technology principles and research progresses of ZFNs, TALENs, CRISPR/Cas9 are introduced.The significant progresses of these emerging technologies achieved in immunodeficient ani-mal models are also elaborated, including KO Rag1/Rag2 rabbit, KO Rag1/Rag2 pig, KO IL2rg pig, KO Ppar-g/Rag1 monkey, and so on.In addition to being models for researching SCID-related diseases in humans, and evaluating the effica-cy and safety of stem-cell engraftment, these models may be also useful to develop surgical procedures for placement of grafts before clinical trials in humans, to produce humanized animals and bridge the gap between laboratory animal and medicical research.The immunodeficient animal models described here represent a step toward the comprehensive evalua-tion of preclinical cellular regenerative strategies.