Nanomolar cellular antisense activity of peptide nucleic acid (PNA) cholic acid ("umbrella") and cholesterol conjugates delivered by cationic lipids.
Nanomolar cellular antisense activity of peptide nucleic acid (PNA) cholic acid ("umbrella") and cholesterol conjugates delivered by cationic lipids.
复制标题
肽核酸 (PNA) 胆酸（“伞”）和阳离子脂质递送的胆固醇缀合物的纳摩尔细胞反义活性。
作者：
Takehiko Shiraishi;P. Nielsen
有限的细胞摄取和较低的生物利用度限制了PNA作为反义/抗基因药物在培养细胞中的广泛应用，尤其是在体内的应用。我们现在报道了胆固醇和胆酸(“伞”)衍生物的剪接校正反义PNA寡聚体的合成及其在培养的HeLa pLuc705细胞中的反义活性。虽然单独的结合物在1μM以下几乎没有活性，但当用阳离子脂质转染剂(LipofetAMINE2000)递送时，它们的活性显著提高。特别是，通过琥珀酸半酯连接到胆固醇或胆酸的PNA表现出低的纳摩尔活性(EC(50)∼25 nM)。由于错配的PNA偶联物没有显示任何显著的反义活性，因此保留了良好的序列特异性。此外，我们还表明，增加转染量提高了转染率，这表明PNA/脂质复合体在细胞表面的积累(凝聚)是摄取机制的一部分。这些结果为PNA低聚物的细胞输送提供了一种新的、简单的方法，特别是使用PNA-胆酸结合物，与PNA-胆固醇结合物相比，PNA-胆酸结合物表现出足够的水溶性。结果还质疑了使用胆酸“伞形”衍生物作为反义低聚物的一种输送方式的普遍性。
Limited cellular uptake and low bioavailability of peptide nucleic acids (PNAs) have restricted widespread use of PNAs as antisense/antigene agents for cells in culture and not least for in vivo applications. We now report the synthesis and cellular antisense activity in cultured HeLa pLuc705 cells of cholesterol and cholic acid ("umbrella") derivatives of splice correction antisense PNA oligomers. While the conjugates alone were practically inactive up to 1 μM, their activity was dramatically improved when delivered by a cationic lipid transfection agent (LipofectAMINE2000). In particular, PNAs, conjugated to cholesterol through an ester hemisuccinate linker or to cholic acid, exhibited low nanomolar activity (EC(50) ∼ 25 nM). Excellent sequence specificity was retained, as mismatch PNA conjugates did not show any significant antisense activity. Furthermore, we show that increasing the transfection volume improved transfection efficiency, suggesting that accumulation (condensation) of the PNA/lipid complex on the cellular surface is part of the uptake mechanism. These results provide a novel, simple method for very efficient cellular delivery of PNA oligomers, especially using PNA-cholic acid conjugates which, in contrast to PNA-cholesterol conjugates, exhibit sufficient water solubility. The results also question the generality of using cholic acid "umbrella" derivatives as a delivery modality for antisense oligomers.
影响因子：
2.9
作者：
Kang, SH;Cho, MJ;Kole, R
通讯作者：
Kole, R