Targeted genome editing is a continually evolving technology employing programmable nucleases to specifically change, insert, or remove a genomic sequence of interest. These advanced molecular tools include meganucleases, zinc finger nucleases, transcription activator-like effector nucleases and RNA-guided engineered nucleases (RGENs), which create double-strand breaks at specific target sites in the genome, and repair DNA either by homologous recombination in the presence of donor DNA or via the error-prone non-homologous end-joining mechanism. A recently discovered group of RGENs known as CRISPR/Cas9 gene-editing systems allowed precise genome manipulation revealing a causal association between disease genotype and phenotype, without the need for the reengineering of the specific enzyme when targeting different sequences. CRISPR/Cas9 has been successfully employed as an ex vivo gene-editing tool in embryonic stem cells and patient-derived stem cells to understand pancreatic beta-cell development and function. RNA-guided nucleases also open the way for the generation of novel animal models for diabetes and allow testing the efficiency of various therapeutic approaches in diabetes, as summarized and exemplified in this manuscript.

中文翻译：

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通过用于胰岛素基因治疗的可编程核酸酶进行基因组工程和疾病建模；CRISPR/Cas9 技术的承诺。

靶向基因组编辑是一项不断发展的技术，它采用可编程核酸酶来特异性地改变、插入或移除感兴趣的基因组序列。这些先进的分子工具包括大范围核酸酶、锌指核酸酶、转录激活因子样效应核酸酶和 RNA 引导的工程核酸酶 (RGEN)，它们在基因组的特定目标位点产生双链断裂，并通过同源重组修复 DNA。供体 DNA 的存在或通过容易出错的非同源末端连接机制。最近发现的一组被称为 CRISPR/Cas9 基因编辑系统的 RGEN 允许精确的基因组操作，揭示疾病基因型和表型之间的因果关系，而无需在靶向不同序列时重新设计特定的酶。CRISPR/Cas9 已成功用作胚胎干细胞和患者来源的干细胞中的离体基因编辑工具，以了解胰腺 β 细胞的发育和功能。RNA 引导的核酸酶还为生成新的糖尿病动物模型开辟了道路，并允许测试各种糖尿病治疗方法的效率，如本手稿中所总结和例证的。