基因人源化苯丙酮尿症小鼠模型作为受精卵人类基因组编辑的测试工具。

IF 4.2 2区 医学 Q1 ENDOCRINOLOGY & METABOLISM Journal of Inherited Metabolic Disease Pub Date : 2024-10-08 DOI:10.1002/jimd.12803
Atsumi Tsuji-Hosokawa, Iku Tsuchiya, Kie Shimizu, Miho Terao, Mito Yasuhara, Natsuho Miyamoto, Saki Kikuchi, Yuya Ogawa, Kazuaki Nakamura, Yoichi Matsubara, Shuji Takada
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引用次数: 0

摘要

靶向基因组编辑取得了重大进展;然而,安全性和伦理问题尚未完全阐明,因此对该技术进行了严格控制。我们利用苯丙酮尿症(PKU)小鼠模型对基因人源化小鼠配子的基因组编辑工具进行了测试,以深入了解人类胚胎的基因组编辑。人类 PKU 小鼠模型 PahhR111X 小鼠已经产生。在 PahhR111X 小鼠中,Pah 的外显子 3 和内含子 3 之间的连接区被 120 bp 的相应人类 PAH 序列取代,其中包括致病性常见变异 c.331C > T。PahhR111X 小鼠成功地重现了 PKU 表型,并表现出认知功能障碍和类似抑郁的行为,这在人类 PKU 患者身上也能观察到。利用靶向人类序列的 sgRNA 对 PahhR111X 小鼠的受精卵进行了基因组编辑。具有校正等位基因的小鼠血清苯丙氨酸水平正常。通过基因组编辑,我们验证了 sgRNA 的实用性。基因人源化小鼠模型揭示了表型的恢复,从而表明对致病变体进行种系基因组编辑对于单基因遗传疾病是可行的;然而,该工具仍存在一些问题,包括其效率和准确性。这项利用基因人化小鼠模型的基因组编辑方案将为改善目前的问题提供启示,并有助于建立可遗传的人类基因组编辑方案。
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Genetically humanized phenylketonuria mouse model as a testing tool for human genome editing in fertilized eggs.

Targeted genome editing has made significant advancements; however, safety and ethical issues have not been fully elucidated, resulting in strict control of the technique. We tested genome editing tools on gametes from a genetically humanized mouse model using a phenylketonuria (PKU) mouse model to gain insights into genome editing in human embryos. The human PKU mouse model PahhR111X mice was generated. The junctional region between exon 3 and intron 3 of Pah was replaced with a 120 bp corresponding human PAH sequence, including the pathogenic common variant c.331C > T in PahhR111X mice. PahhR111X mice successfully recapitulated the PKU phenotype and showed cognitive dysfunction and depressive-like behavior, which are observed in human patients with PKU. Genome editing was applied to fertilized eggs of PahhR111X mice utilizing sgRNA that targets the human sequence. Mice with the corrected allele exhibited normal serum phenylalanine levels. Through genome editing, we validated the utility of sgRNA. The genetically humanized mouse model suggested that germ-line genome editing of the pathogenic variant may be feasible for monogenic disorders by revealing the recovery of the phenotype; however, there are remaining issues with the tool, including its efficiency and accuracy. This genome editing protocol using a genetically humanized mouse model will provide insights for improving current issues and contribute to the establishment of heritable human genome editing protocols.

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来源期刊
Journal of Inherited Metabolic Disease
Journal of Inherited Metabolic Disease 医学-内分泌学与代谢
CiteScore
9.50
自引率
7.10%
发文量
117
审稿时长
4-8 weeks
期刊介绍: The Journal of Inherited Metabolic Disease (JIMD) is the official journal of the Society for the Study of Inborn Errors of Metabolism (SSIEM). By enhancing communication between workers in the field throughout the world, the JIMD aims to improve the management and understanding of inherited metabolic disorders. It publishes results of original research and new or important observations pertaining to any aspect of inherited metabolic disease in humans and higher animals. This includes clinical (medical, dental and veterinary), biochemical, genetic (including cytogenetic, molecular and population genetic), experimental (including cell biological), methodological, theoretical, epidemiological, ethical and counselling aspects. The JIMD also reviews important new developments or controversial issues relating to metabolic disorders and publishes reviews and short reports arising from the Society''s annual symposia. A distinction is made between peer-reviewed scientific material that is selected because of its significance for other professionals in the field and non-peer- reviewed material that aims to be important, controversial, interesting or entertaining (“Extras”).
期刊最新文献
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