美国纽约大学朗贡医疗中心Jef D. Boeke.研究团队近日取得一项新成果。经过不懈努力，他们完成了小鼠基因组重建和对三个重要疾病位点的改造。相关论文发表在2023年11月1日出版的《自然》杂志上。
现有的方法对运送DNA的大小和效率存在局限，妨碍了高信息量模型的常规创建，研究人员称之为基因组改写和定制的基因工程小鼠模型（GEMMs）（GREAT-GEMMs）。
研究人员研发了"哺乳动物抗生素耐药性标记逐步转换整合"（mSwAP-In）技术，这是一种在小鼠胚胎干细胞中高效重建基因组的方法。研究人员展示了使用mSwAP-In对长115k的Trp53基因座进行迭代基因组重建，以及使用116kb和180kb的人类ACE2基因座对小鼠进行人源化。ACE2模型再现了人类ACE2的表达模式和剪接，与现有的K18-hACE2模型相比，ACE2模型在感染SARS-CoV-2时的症状更轻，因此代表了一种更类似人类的感染模型。最后，研究通过在ACE2 GREAT-GEMM中对小鼠Tmprss2进行双叶人源化，演示了序列基因组编写，突出了mSwAP-In在基因组编写中的多功能性。
据了解，GEMMs有助于人们了解人类相关疾病的病理并开发新的疗法，然而在小鼠身上完全再现人类疾病却极具挑战性。基因组学的进步凸显了非编码调控基因组序列的重要性，这些序列参与调控与许多人类疾病有关基因的时空表达模式和剪接。囊括广泛的调控基因组区域需要大规模基因组工程，这将提高疾病建模的质量。
附：英文原文
Title: Mouse genome rewriting and tailoring of three important disease loci
Author: Zhang, Weimin, Golynker, Ilona, Brosh, Ran, Fajardo, Alvaro, Zhu, Yinan, Wudzinska, Aleksandra M., Ordoez, Raquel, Ribeiro-dos-Santos, Andr M., Carrau, Lucia, Damani-Yokota, Payal, Yeung, Stephen T., Khairallah, Camille, Vela Gartner, Antonio, Chalhoub, Noor, Huang, Emily, Ashe, Hannah J., Khanna, Kamal M., Maurano, Matthew T., Kim, Sang Yong, tenOever, Benjamin R., Boeke, Jef D.
Issue&Volume: 2023-11-01
Abstract: Genetically engineered mouse models (GEMMs) help us to understand human pathologies and develop new therapies, yet faithfully recapitulating human diseases in mice is challenging. Advances in genomics have highlighted the importance of non-coding regulatory genome sequences, which control spatiotemporal gene expression patterns and splicing in many human diseases1,2. Including regulatory extensive genomic regions, which requires large-scale genome engineering, should enhance the quality of disease modelling. Existing methods set limits on the size and efficiency of DNA delivery, hampering the routine creation of highly informative models that we call genomically rewritten and tailored GEMMs (GREAT-GEMMs). Here we describe ‘mammalian switching antibiotic resistance markers progressively for integration’ (mSwAP-In), a method for efficient genome rewriting in mouse embryonic stem cells. We demonstrate the use of mSwAP-In for iterative genome rewriting of up to 115kb of a tailored Trp53 locus, as well as for humanization of mice using 116kb and 180kb human ACE2 loci. The ACE2 model recapitulated human ACE2 expression patterns and splicing, and notably, presented milder symptoms when challenged with SARS-CoV-2 compared with the existing K18-hACE2 model, thus representing a more human-like model of infection. Finally, we demonstrated serial genome writing by humanizing mouse Tmprss2 biallelically in the ACE2 GREAT-GEMM, highlighting the versatility of mSwAP-In in genome writing.
DOI: 10.1038/s41586-023-06675-4
Source: https://www.nature.com/articles/s41586-023-06675-4
Nature：《自然》，创刊于1869年。隶属于施普林格·自然出版集团，最新IF：69.504
官方网址：http://www.nature.com/
投稿链接：http://www.nature.com/authors/submit_manuscript.html
本期文章：《自然》：Online/在线发表
