Aptamer-guided gene targeting in yeast and human cells.
Aptamer-guided gene targeting in yeast and human cells.
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DOI：
10.1093/nar/gku101
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Storici F
中科院分区：
文献类型：
--
作者：
Ruff P;Koh KD;Keskin H;Pai RB;Storici F
基因靶向是通过同源重组（HR）修饰内源DNA序列在其基因组位置的遗传技术，并且可用于功能分析和基因治疗应用。HR在大多数生物体和细胞类型（包括哺乳动物细胞）中效率低下，通常限制了基因靶向的有效性。因此，提高人力资源效率仍然是DNA编辑的主要挑战。在这里，我们提出了一个新的概念，基因校正的基础上开发的DNA适体能够结合到一个位点特异性的DNA结合蛋白，以促进供体分子和所需的靶位点之间的同源遗传信息的交换（适体引导的基因靶向）。我们选择了I-SceI核酸内切酶的DNA适体。将含有I-SceI适体序列的双功能寡核苷酸设计为较长单链DNA分子的一部分，所述较长单链DNA分子含有具有同源性的区域以修复I-SceI产生的双链断裂并校正破坏的基因。含I-SceI适体的寡核苷酸在酵母酿酒酵母中刺激基因靶向高达32倍，在人类细胞中高达16倍。本工作为提高基因打靶效率提供了一个新的概念和研究方向，并为今后利用适体进行基因打靶的研究奠定了基础。
Gene targeting is a genetic technique to modify an endogenous DNA sequence in its genomic location via homologous recombination (HR) and is useful both for functional analysis and gene therapy applications. HR is inefficient in most organisms and cell types, including mammalian cells, often limiting the effectiveness of gene targeting. Therefore, increasing HR efficiency remains a major challenge to DNA editing. Here, we present a new concept for gene correction based on the development of DNA aptamers capable of binding to a site-specific DNA binding protein to facilitate the exchange of homologous genetic information between a donor molecule and the desired target locus (aptamer-guided gene targeting). We selected DNA aptamers to the I-SceI endonuclease. Bifunctional oligonucleotides containing an I-SceI aptamer sequence were designed as part of a longer single-stranded DNA molecule that contained a region with homology to repair an I-SceI generated double-strand break and correct a disrupted gene. The I-SceI aptamer-containing oligonucleotides stimulated gene targeting up to 32-fold in yeast Saccharomyces cerevisiae and up to 16-fold in human cells. This work provides a novel concept and research direction to increase gene targeting efficiency and lays the groundwork for future studies using aptamers for gene targeting.
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影响因子：
3.9
作者：
Cowperthwaite, Matthew C.;Ellington, Andrew D.
通讯作者：
Ellington, Andrew D.
DOI：
10.3390/molecules14041353
发表时间：
2009-03-27
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Pan W;Clawson GA
通讯作者：
Clawson GA
DOI：
10.1073/pnas.052545899
发表时间：
2002-03-19
影响因子：
11.1
作者：
Allen, C;Kurimasa, A;Nickoloff, JA
通讯作者：
Nickoloff, JA
影响因子：
7.4
作者：
Berezovski, M;Krylov, SN
通讯作者：
Krylov, SN
影响因子：
5.6
作者：
Niu, Yan;Tenney, Kristen;Gimble, Frederick S.
通讯作者：
Gimble, Frederick S.