Chimeric RNA/DNA oligonucleotide-based gene therapy.
Chimeric RNA/DNA oligonucleotide-based gene therapy.
复制标题
基于嵌合 RNA/DNA 寡核苷酸的基因治疗。
DOI：
10.1046/j.1523-1755.2002.0610s1047.x
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发表时间：
2002
影响因子：
19.6
通讯作者：
Lien,Yeong-HauH
中科院分区：
文献类型：
--
作者：
Lai,Li-Wen;Lien,Yeong-HauH
基于嵌合RNA/DNA寡核苷酸的基因治疗背景嵌合RNA/DNA寡核苷酸作为一种潜在的基因治疗策略，已被证明在几种遗传病模型中诱导点突变的定点特异性纠正。嵌合的RNA/DNA寡核苷酸，除了中心的错配外，与突变位点两侧的25到30个基因组DNA残基互补，通过不同的途径和递送工具输送到不同的组织和器官。用等位基因特异性聚合酶链式反应、限制性内切酶消化、菌落提升分析、测序、Northern和Western印迹分析、酶活性测定、免疫组织化学染色和功能研究等方法对突变在基因和表型水平上的校正进行了评估。与同源重组频率(10−5~10−6)相比，转化率高出好几倍。某些研究表明，这种表型变化持续了一年以上。结论基于嵌合寡核苷酸的基因治疗有望成为治疗遗传病的有效手段。它可以在适当的细胞或组织中提供校正基因的永久表达和正常调节。为了提高这种方法的有效性和安全性，有必要进一步阐明基于RNA/DNA寡核苷酸的嵌合基因治疗的机制。
Chimeric RNA/DNA oligonucleotide-based gene therapyBackgroundChimeric RNA/DNA oligonucleotides, emerging as a potential strategy for gene therapy, have been shown to induce site-specific correction of point mutations in several genetic disease models.MethodsSix recent studies of chimeric RNA/DNA oligonucleotide-based gene therapy in genetic disease models are reviewed. Chimeric RNA/DNA oligonucleotides, complementary to 25 to 30 residues of genomic DNA flanking the mutation site with the exception of a mismatch in the center, were delivered via different routes and delivery vehicles to target different tissues and organs. Corrections of the mutation at genotypic and phenotypic levels were assessed using various methods, including allele-specific polymerase chain reaction assay, restriction enzyme digestion, colony-lifting assays, sequencing, Northern and Western blot analyses, enzyme activity assay, immunohistochemical staining, and functional studies.ResultsThe gene correction frequency varied, ranging from less than 1% to more than 40%. This represented several magnitudes higher conversion rate compared with homologous recombination frequency, which is in the range of 10−5to 10−6. The resulting phenotype changes lasted longer than one year in some studies.ConclusionChimeric RNA/DNA oligonucleotide-based gene therapy has the potential to develop into powerful therapeutic modality for genetic diseases. It can offer permanent expression and normal regulation of corrected genes in appropriate cells or tissues. Further efforts to elucidate the mechanisms of chimeric RNA/DNA oligonucleotide-based gene therapy are warranted in order to increase the efficacy and safety of this method.