Hammerhead ribozymes targeted to the FBN1 mRNA can discriminate a single base mismatch between ribozyme and target.
Hammerhead ribozymes targeted to the FBN1 mRNA can discriminate a single base mismatch between ribozyme and target.
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靶向 FBN1 mRNA 的锤头核酶可以区分核酶和靶标之间的单碱基错配。
DOI：
10.1006/bbrc.1998.9241
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发表时间：
1998
影响因子：
3.1
通讯作者：
Michael W. Kilpatrick
中科院分区：
文献类型：
--
作者：
Leo Phylactou;Leo Phylactou;Petros Tsipouras;Michael W. Kilpatrick
锤头状核酶是一种催化反式作用的RNA分子，核酶和底物是两种不同的具有互补区的寡核苷酸。纤维蛋白-1(FBN1)基因的突变会导致马凡综合征。大多数突变是单碱基改变，其中许多是通过显性-负机制发挥作用。以前我们已经证明，针对FBN1 mRNA的反义锤头状核酶可以减少纤维蛋白在培养的成纤维细胞细胞外基质中的沉积，这表明利用核酶下调突变蛋白的产生，从而恢复正常的纤维蛋白功能是可能的。这可能是通过突变产生一个正常等位基因中不存在的核酶切割位点来实现的，然而这可能会限制可以靶向的突变的数量。或者，也可以通过核酶结合臂来靶向突变的等位基因。为了确定核酶通过后一种方法优先靶向突变的FBN1等位基因的可能性，研究了锤头状核酶螺旋I错配对纤维蛋白(FBN1)mRNA切割的影响。在体外和体内，单碱基错配都显著降低了核酶的切割效率。完全匹配核酶和错配核酶的区别随着螺旋I长度的不同而不同，这种区别在较短螺旋的核酶中更为明显。这些数据表明，设计锤头状核酶应该是可能的，即使突变不会产生核酶切割位点，也可以区分密切相关的(突变的和正常的)目标RNA，差异小到单个核苷酸。
Hammerhead ribozymes are catalytic RNA molecules that can act in trans, with ribozyme and substrate being two different oligoribonucleotides with regions of complementarity. Mutations in the gene for fibrillin-1 (FBN1) cause Marfan syndrome. The majority of mutations are single-base changes, many of which exert their effect via a dominant-negative mechanism. Previously we have shown that an antisense hammerhead ribozyme, targeted to the FBN1 mRNA can reduce deposition of fibrillin to the extracellular matrix of cultured fibroblasts, suggesting it may be possible to utilize ribozymes to down regulate the production of mutant protein and thus restore normal fibrillin function. This might be achieved by the mutation creating a ribozyme cleavage site that is not present in the normal allele, however this is likely to limit the number of mutations that could be targeted. Alternatively, it might be possible to target the mutant allele via the ribozyme binding arms. To determine the potential of ribozymes to preferentially target mutant FBN1 alleles via the latter approach, the effect of mismatches in helix I of a hammerhead ribozyme, on the cleavage of fibrillin (FBN1) mRNA was investigated. A single base mismatch significantly reduced ribozyme cleavage efficiency both in vitro and in vivo. The discrimination between fully-matched and mismatched ribozyme varied with the length of helix I, with the discrimination being more pronounced in ribozymes with a shorter helix. These data suggest that it should be possible to design hammerhead ribozymes that can discriminate between closely related (mutant and normal) target RNAs varying in as little as a single nucleotide, even if the mutation does not create a ribozyme cleavage site.
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影响因子：
9.8
作者：
M. Godfrey;Natalie Vandemark;Mei Wang;M. Velinov;D. Wargowski;P. Tsipouras;Jenny Han;J. Becker;W. L. Robertson;S. Droste;V. Rao
通讯作者：
M. Godfrey;Natalie Vandemark;Mei Wang;M. Velinov;D. Wargowski;P. Tsipouras;Jenny Han;J. Becker;W. L. Robertson;S. Droste;V. Rao
影响因子：
56.9
作者：
SARVER, N;CANTIN, EM;ROSSI, JJ
通讯作者：
ROSSI, JJ
影响因子：
4.4
作者：
DIETZ, HC;MCINTOSH, I;FRANCOMANO, CA
通讯作者：
FRANCOMANO, CA
锤头核酶底物识别螺旋中碱基错配对结合和催化的影响。
影响因子：
14.9
作者：
Werner,M;Uhlenbeck,OC
通讯作者：
Uhlenbeck,OC
通过抗癌基因核酶抑制黑色素瘤细胞的恶性表型。
DOI：
10.1111/1523-1747.ep12340688
发表时间：
1996
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
Ohta,Y;Kijima,H;Kashani-Sabet,M;Scanlon,KJ
通讯作者：
Scanlon,KJ