Triplex-induced recombination and repair in the pyrimidine motif.
Triplex-induced recombination and repair in the pyrimidine motif.
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嘧啶基序中三环诱导的重组和修复。
DOI：
10.1093/nar/gki659
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发表时间：
2005
影响因子：
14.9
通讯作者：
Glazer, PM
中科院分区：
文献类型：
--
作者：
Kalish, JM;Seidman, MM;Weeks, DL;Glazer, PM
三链形成寡核苷酸(TFOS)以序列特异性的方式与DNA结合在多嘌呤/多嘧啶位点，并介导靶向基因组修饰。三链体是由与双链嘌呤链(嘧啶，平行基序)平行结合的嘧啶TFOS或以反平行取向(嘌呤，反平行基序)结合的嘌呤TFOS形成的。嘌呤和嘧啶TFOS，当连接到补骨脂素时，已被证明在细胞中引导补骨脂素加合物的形成，导致突变或重组。然而，只有嘌呤TFOS被证明可以在不需要靶向DNA加合物的情况下介导基因组修饰。在这项工作中，我们报告了一系列的嘧啶TFOS，经过选择的化学修饰，在没有任何DNA反应结合物的情况下，在哺乳动物细胞中诱导两个不同的异构体靶点修复和重组的能力。我们发现，在生理条件下，与未修饰的→TFOS相比，含有N3‘DNAP5’磷酰胺(酰胺)、5-(1-丙炔基)-2‘-脱氧尿苷(PDU)、2’-O-甲基核糖(2‘-O-Me)、2’-O-(2-氨乙基)-核糖、2‘-O，4’-亚甲基桥联或锁定核酸的TFOS显著增加了非共价三链的形成。然而，在这些修饰的TFOS中，只有酰胺和PDU修饰的TFOS介导了细胞中诱导的重组并刺激了细胞提取物中的修复，其水平与类似检测中的嘌呤TFOS相当。这些结果表明，酰亚胺和PDU修饰的TFOS可以作为试剂来刺激位点特异性基因打靶，而不需要与DNA反应分子结合。通过展示适当修饰的嘧啶TFOS诱导修复和重组的可能性，这项工作扩大了三链介导的基因靶向的可用选择。
Triplex-forming oligonucleotides (TFOs) bind DNA in a sequence-specific manner at polypurine/polypyrimidine sites and mediate targeted genome modification. Triplexes are formed by either pyrimidine TFOs, which bind parallel to the purine strand of the duplex (pyrimidine, parallel motif), or purine TFOs, which bind in an anti-parallel orientation (purine, anti-parallel motif). Both purine and pyrimidine TFOs, when linked to psoralen, have been shown to direct psoralen adduct formation in cells, leading to mutagenesis or recombination. However, only purine TFOs have been shown to mediate genome modification without the need for a targeted DNA-adduct. In this work, we report the ability of a series of pyrimidine TFOs, with selected chemical modifications, to induce repair and recombination in two distinct episomal targets in mammalian cells in the absence of any DNA-reactive conjugate. We find that TFOs containing N3′→P5′ phosphoramidate (amidate), 5-(1-propynyl)-2′-deoxyuridine (pdU), 2′-O-methyl-ribose (2′-O-Me), 2′-O-(2-aminoethyl)-ribose, or 2′-O, 4′-C-methylene bridged or locked nucleic acid (LNA)-modified nucleotides show substantially increased formation of non-covalent triplexes under physiological conditions compared with unmodified DNA TFOs. However, of these modified TFOs, only the amidate and pdU-modified TFOs mediate induced recombination in cells and stimulate repair in cell extracts, at levels comparable to those seen with purine TFOs in similar assays. These results show that amidate and pdU-modified TFOs can be used as reagents to stimulate site-specific gene targeting without the need for conjugation to DNA-reactive molecules. By demonstrating the potential for induced repair and recombination with appropriately modified pyrimidine TFOs, this work expands the options available for triplex-mediated gene targeting.
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影响因子：
14.9
作者：
FRANCOIS, JC;SAISONBEHMOARAS, T;HELENE, C
通讯作者：
HELENE, C
DOI：
10.1073/pnas.94.1.79
发表时间：
1997-01-07
影响因子：
11.1
作者：
Giovannangeli, C;Diviacco, S;Helene, C
通讯作者：
Helene, C
DOI：
10.1073/pnas.92.13.5798
发表时间：
1995-06-20
影响因子：
11.1
作者：
GRYAZNOV, SM;LLOYD, DH;WILSON, WD
通讯作者：
WILSON, WD
影响因子：
14.9
作者：
Holland, JA;Hoffman, DW
通讯作者：
Hoffman, DW
DOI：
10.1006/bbrc.2000.2438
发表时间：
2000-04-13
影响因子：
3.1
作者：
Lacroix, L;Arimondo, PB;Mergny, JL
通讯作者：
Mergny, JL