Redirecting splicing with bifunctional oligonucleotides.
Redirecting splicing with bifunctional oligonucleotides.
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用双功能寡核苷酸重定向剪接。
DOI：
10.1093/nar/gkt1287
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Chabot B
中科院分区：
文献类型：
--
作者：
Brosseau JP;Lucier JF;Lamarche AA;Shkreta L;Gendron D;Lapointe E;Thibault P;Paquet E;Perreault JP;Abou Elela S;Chabot B
选择性剪接的异位调节剂是研究剪接变异体功能和纠正导致人类疾病的剪接错误事件的重要工具。这种调节剂可以是双功能寡核苷酸，由决定靶标特异性的反义部分和募集影响剪接位点选择的蛋白质或RNA/蛋白复合物的非杂交尾部组成（分别为靶向寡核苷酸剪接沉默者和靶向寡核苷酸剪接增强者的TOSS和TOES）。TOSS和TOES的使用仅限于少数目标。为了推广TOSS的适用性并证明其健壮性，我们在50多个备选剪接事件上对该方法进行了测试。此外，我们还开发了一种算法，可以设计出成功率为80%的主动TOSS。为了产生能够刺激剪接的双功能寡核苷酸，我们观察到TDP-43的结合位点可以刺激剪接，当插入5 '剪接位点下游时，可以改善U1 snRNP的结合。一个旨在招募TDP-43的TOES改善了SMN2中外显子7的内含性。总的来说，我们的研究表明，双功能寡核苷酸可以重定向各种基因的剪接，证明将它们包含在旨在改变剪接变异体产生的分子库中是合理的。
Ectopic modulators of alternative splicing are important tools to study the function of splice variants and for correcting mis-splicing events that cause human diseases. Such modulators can be bifunctional oligonucleotides made of an antisense portion that determines target specificity, and a non-hybridizing tail that recruits proteins or RNA/protein complexes that affect splice site selection (TOSS and TOES, respectively, for targeted oligonucleotide silencer of splicing and targeted oligonucleotide enhancer of splicing). The use of TOSS and TOES has been restricted to a handful of targets. To generalize the applicability and demonstrate the robustness of TOSS, we have tested this approach on more than 50 alternative splicing events. Moreover, we have developed an algorithm that can design active TOSS with a success rate of 80%. To produce bifunctional oligonucleotides capable of stimulating splicing, we built on the observation that binding sites for TDP-43 can stimulate splicing and improve U1 snRNP binding when inserted downstream from 5′ splice sites. A TOES designed to recruit TDP-43 improved exon 7 inclusion in SMN2. Overall, our study shows that bifunctional oligonucleotides can redirect splicing on a variety of genes, justifying their inclusion in the molecular arsenal that aims to alter the production of splice variants.
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影响因子：
46.9
作者：
Judge, AD;Sood, V;MacLachlan, I
通讯作者：
MacLachlan, I
影响因子：
3.5
作者：
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作者：
Hua, Yimin;Vickers, Timothy A;Baker, Brenda F;Bennett, C Frank;Krainer, Adrian R
通讯作者：
Krainer, Adrian R
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通讯作者：
Khvorova, Anastasia
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10.5
作者：
Hua, Yimin;Sahashi, Kentaro;Krainer, Adrian R.
通讯作者：
Krainer, Adrian R.