Allele-Specific Silencing by RNA Interference
Allele-Specific Silencing by RNA Interference
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DOI：
10.1007/978-1-60761-588-0_4
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发表时间：
2010-01-01
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影响因子：
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通讯作者：
Hohjoh, Hirohiko
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作者：
Hohjoh, Hirohiko
通过RNA干扰（RNAi）的等位基因特异性基因沉默在治疗上可用于特异性抑制疾病相关等位基因的表达而不抑制相应野生型等位基因的表达。为了实现这样的等位基因特异性RNAi（ASP-RNAi），赋予ASP-RNAi的小干扰RNA（siRNA）双链体的设计和评估是至关重要的;然而，这也是困难的。我们已经开发了一种具有报告等位基因的测定系统，所述报告等位基因编码Photinus和Renilla荧光素酶基因，所述荧光素酶基因在其3 '-非翻译区中携带突变型和野生型等位基因序列。该测定系统允许我们以定性和定量的方式同时评估针对靶突变等位基因的等位基因特异性沉默以及针对相应野生型等位基因的脱靶沉默的设计的siRNA以及短发夹RNA。
Allele-specific gene silencing by RNA interference (RNAi) is therapeutically useful for specifically inhibiting the expression of disease-associated alleles without suppressing the expression of corresponding wild-type alleles. To realize such allele-specific RNAi (ASP-RNAi), the design and assessment of small interfering RNA (siRNA) duplexes conferring ASP-RNAi is vital; however, it is also difficult. We have developed an assay system with reporter alleles that encode the Photinus and Renilla luciferase genes carrying mutant and wild-type allelic sequences in their 3'-untranslated regions. The assay system allows us to evaluate designed siRNAs and also short hairpin RNAs for allele-specific silencing against target mutant alleles as well as off-target silencing against corresponding wild-type alleles simultaneously in a qualitative and quantitative manner.