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准一种致病密码外基因,驱使HLRCC诱导外基因跳转

Siddhardha S Maligireddy¹, Mariana D Mandler¹, Judith C Lunger¹

¹Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.

Molecular therapy. Nucleic acids

|September 2, 2025

概括

对遗传性菌瘤和细胞癌 (HLRCC) 进行了研究,以恢复 fumarate hydratase (FH) 的表达. 使用CRISPR-Cas9和反感性寡核酸 (ASOs) 的拼接调制成功排除了一个神秘的外基子,提供了治疗潜力.

科学领域:

遗传学
分子生物学
癌症学

背景情况:

遗传性骨髓瘤和细胞癌 (HLRCC) 是一种遗传性癌症综合征,由烟酸酶 (FH) 基因的功能丧失突变引起.
在FH基因的第9个内核中,一种特定的致病变体破坏了正常的拼接,导致一个神秘的外核和一个过早的终结,最终导致FH缺乏.
恢复功能性FH蛋白表达对于具有这种特定拼接缺陷的HLRC患者的治疗干预至关重要.

研究的目的:

确定和测试调节FH基因拼接的策略,以排除神秘的外基因.
评估基因组编辑和反意义寡核酸 (ASOs) 在纠正拼接缺陷方面的有效性.
评估拼接调制作为HLRCC治疗方法的潜力.

主要方法:

开发一个小基因GFP报告系统来模拟FH拼接缺陷.
应用CRISPR-Cas9基因组编辑来准和纠正拼接异常.
使用反感性寡核酸 (ASOs) 来促进密码外基的跳转.
在患者衍生的纤维细胞中进行验证,以证明FH mRNA异形的异位特异调节.

主要成果:

无论是CRISPR-Cas9还是ASO都成功地诱导了记者细胞系中的神秘外基因跳转.
在患者细胞中,ASO证明了由参考和变异性等位基因衍生的FHmRNA异位基因平衡的能力.
开发的报告系统有效地回顾了HLRCC患者观察到的拼接缺陷.

结论:

包括ASO和基因组编辑在内的拼接调节策略显示出对纠正HLRCC中的FH拼接缺陷的希望.
对非编码FH突变引起的HLRCC而言,反意义寡核酸是一种潜在的治疗途径.
这些发现支持开发针对性疗法,旨在恢复HLRCC的功能.

关键词:

其他国家在CRISPR-Cas9中美国美国 MT:氧核酸:治疗方法和应用接合方式

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