Strategems in vitro for gene therapies directed to dominant mutations
Strategems in vitro for gene therapies directed to dominant mutations
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DOI：
10.1093/hmg/6.9.1415
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发表时间：
1997-09-01
影响因子：
3.5
通讯作者：
Farrar, GJ
中科院分区：
文献类型：
--
作者：
MillingtonWard, S;ONeill, B;Farrar, GJ
与设计用于常染色体显性疾病的基因疗法相关的主要困难是在此类病症中经常遇到的巨大的基因内异质性。为了克服此类困难，我们设计并在体外评估了三种策略，其避免了靶向个体突变用于遗传抑制的要求。正常和突变等位基因被转录但未翻译的转录物区域（UTR）中的靶向序列抑制，使得能够引入具有正确编码序列但改变UTR的替代基因以防止抑制，第二种方法涉及通过利用遗传密码的简并性抑制编码序列并同时引入替代基因。第三种策略利用基因内多态性来特异性抑制疾病等位基因，优点是具有不同疾病突变的患者的比例具有相同的多态性，这些方法提供了比针对单一疾病突变的靶序列更广泛的靶序列选择，并且适用于给定基因内的许多突变，抑制的一般方法可以针对与疾病过程相关的原发性缺陷或继发性效应，例如凋亡，三个一般的方法，针对主要缺陷，规避问题的等位基因遗传异质性进行了探讨，在体外使用锤头设计为靶向来自视紫红质、外周蛋白和胶原1A1和1A2基因的转录物的核酶，广泛的遗传异质性是相关疾病病理学的特征。
A major difficulty associated with the design of gene therapies for autosomal dominant diseases is the immense intragenic heterogeneity often encountered in such conditions, In order to overcome such difficulties we have designed, and evaluated in vitro, three strategies which avoid a requirement to target individual mutations for genetic suppression, In the first, normal and mutant alleles are suppressed by targeting sequences in transcribed but untranslated regions of transcript (UTRs), enabling introduction of a replacement gene with the correct coding sequencing but altered UTRs to prevent suppression, A second approach involves suppression in coding sequence and concurrent introduction of a replacement gene by exploiting the degeneracy of the genetic code, A third strategy utilises intragenic polymorphism to suppress the disease allele specifically, the advantage being that a proportion of patients with different disease mutations have the same polymorphism, These approaches provide a wider choice of target sequence than those directed to single disease mutations and are appropriate for many mutations within a given gene, General methods for suppression may be directed towards the primary defect or a secondary effect associated with the disease process, such as apoptosis, Three general methods targeting the primary defect which circumvent problems of allelic genetic heterogeneity are explored in vitro using hammerhead ribozymes designed to target transcripts from the rhodopsin, peripherin and collagen 1A1 and 1A2 genes, extensive genetic heterogeneity being a feature of associated disease pathologies.