2'-deoxy-2'-fluoro-beta-D-arabinonucleic acid (2'F-ANA) modified oligonucleotides (ON) effect highly efficient, and persistent, gene silencing.
2'-deoxy-2'-fluoro-beta-D-arabinonucleic acid (2'F-ANA) modified oligonucleotides (ON) effect highly efficient, and persistent, gene silencing.
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DOI：
10.1093/nar/gkj455
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发表时间：
2006
影响因子：
14.9
通讯作者：
Gewirtz AM
中科院分区：
文献类型：
--
作者：
Kalota A;Karabon L;Swider CR;Viazovkina E;Elzagheid M;Damha MJ;Gewirtz AM
为了在体内有效，反义寡核苷酸（AS ON）应该是核酸酶抗性的，形成稳定的ON/RNA双链体并支持核糖核酸酶H介导的异源双链体切割，所有这些对细胞功能的非特异性影响都可以忽略不计。我们在此报道，含有2′-脱氧-2 ′-氟-β-d-阿拉伯糖核酸（2′F-ANA）糖修饰的AS ON不仅符合这些标准，而且具有延长细胞内浓度的额外优势，这似乎促进了更长时间的基因沉默。为了证明这一点，我们用完全硫代磷酸化的2′F-ANA-DNA嵌合体（PS-2′FANA-DNA）靶向人白血病细胞中的c-MYB原癌基因mRNA，并比较它们与含有未修饰核苷的AS ON（PS-DNA）的基因沉默效率。当通过核转染递送时，两种类型的化学修饰的ON实现了>90%的c-MYB mRNA和蛋白质表达的敲低，但是PS-2′F-ANA-DNA能够在PS-DNA的20%剂量下实现这一点，并且与PS-AS DNA相反，它们的沉默效果在单次施用后4天后仍然存在。因此，我们的数据表明，PS-2′F-ANA-DNA嵌合体是有效的基因沉默分子，并表明它们可能具有显着的治疗潜力。
To be effective in vivo, antisense oligonucleotides (AS ON) should be nuclease resistant, form stable ON/RNA duplexes and support ribonuclease H mediated heteroduplex cleavage, all with negligible non-specific effects on cell function. We report herein that AS ONs containing a 2′-deoxy-2′-fluoro-β-d-arabinonucleic acid (2′F-ANA) sugar modification not only meet these criteria, but have the added advantage of maintaining high intracellular concentrations for prolonged periods of time which appears to promote longer term gene silencing. To demonstrate this, we targeted the c-MYB protooncogene's mRNA in human leukemia cells with fully phosphorothioated 2′F-ANA–DNA chimeras (PS-2′FANA–DNA) and compared their gene silencing efficiency with AS ON containing unmodified nucleosides (PS-DNA). When delivered by nucleofection, chemically modified ON of both types effected a >90% knockdown of c-MYB mRNA and protein expression, but the PS-2′F-ANA–DNA were able to accomplish this at 20% of the dose of the PS-DNA, and in contrast to the PS-AS DNA, their silencing effect was still present after 4 days after a single administration. Therefore, our data demonstrate that PS-2′F-ANA–DNA chimeras are efficient gene silencing molecules, and suggest that they could have significant therapeutic potential.
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DOI：
10.1073/pnas.94.6.2620
发表时间：
1997-03-18
影响因子：
11.1
作者：
Agrawal, S;Jiang, ZW;Zhang, RW
通讯作者：
Zhang, RW
影响因子：
15
作者：
Damha, MJ;Wilds, CJ;Parniak, MA
通讯作者：
Parniak, MA
影响因子：
2.9
作者：
Lok, CN;Viazovkina, E;Parniak, MA
通讯作者：
Parniak, MA
影响因子：
2.7
作者：
Min, KL;Viazovkina, E;Damha, MJ
通讯作者：
Damha, MJ
影响因子：
14.9
作者：
Denisov, AY;Noronha, AM;Damha, MJ
通讯作者：
Damha, MJ