Specific Inhibition of Influenza Virus RNA Polymerase and Nucleoprotein Gene Expression by Liposomally Encapsulated Antisense Phosphorothioate Oligonucleotides in MDCK Cells
Specific Inhibition of Influenza Virus RNA Polymerase and Nucleoprotein Gene Expression by Liposomally Encapsulated Antisense Phosphorothioate Oligonucleotides in MDCK Cells
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脂质体封装的反义硫代磷酸寡核苷酸对 MDCK 细胞中流感病毒 RNA 聚合酶和核蛋白基因表达的特异性抑制
DOI：
10.1177/095632029800900306
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发表时间：
1998
影响因子：
--
通讯作者：
H. Takaku
中科院分区：
文献类型：
--
作者：
T. Abe;S. Suzuki;T. Hatta;K. Takai;T. Yokota;H. Takaku
我们已经证明，反义硫代磷酸寡核苷酸（S-ODNs）抑制流感病毒在MDCK细胞中的复制。用MTT法检测脂质体包裹和游离的具有4个靶位点（PB 1、PB 2、PA和NP基因）的反义S-ODNs抑制病毒诱导的细胞病变的能力。脂质体包裹的S-ODN与PB 2 AUG起始密码子周围的位点互补，显示出高度的抑制作用。与此相反，脂质体包裹的S-ODN靶向PB 1的抑制作用显著降低，与针对PB 2靶位点的抑制作用相比。脂质体包裹的反义S-ODNs表现出更高的抑制活性比游离的寡核苷酸，并显示序列特异性抑制，而观察到游离的反义S-ODNs抑制病毒吸附到MDCK细胞。寡核苷酸的脂质体制剂促进其从内吞囊泡释放，从而观察到细胞质和核定位。脂质体载体能有效提高反义S-ODNs的活性。有趣的是，脂质体包封的FITC-S-ODN-PB 2-as在MDCK细胞的核区域中积累。而游离反义S-ODNs处理的MDCK细胞胞浆和胞内体内可见微弱荧光。因此，阳离子脂质颗粒可以是适合于体外或体内使用的用于基于阿托伐他汀的治疗剂和转基因的潜在有用的递送载体。
We have demonstrated that antisense phosphorothioate oligonucleotides (S-ODNs) inhibit influenza A virus replication in MDCK cells. Liposomally encapsulated and free antisense S-ODNs with four target sites (PB1, PB2, PA and NP genes) were tested for their abilities to inhibit virus-induced cytopathogenic effects in a MTT assay using MDCK cells. The liposomally encapsulated S-ODN complementary to the site around the PB2 AUG initiation codon showed highly inhibitory effects. In contrast, the inhibitory effect of the liposomally encapsulated S-ODN targeted to PB1 was considerably decreased in comparison with that directed to the PB2 target site. The liposomally encapsulated antisense S-ODNs exhibited higher inhibitory activities than the free oligonucleotides, and showed sequence-specific inhibition, whereas free antisense S-ODNs were observed to inhibit viral adsorption to MDCK cells. Liposomal preparations of oligonucleotides facilitated their release from endocytic vesicles, and thus cytoplasmic and nuclear localization was observed. The activities of the antisense S-ODNs were effectively enhanced by using the liposomal carrier. Interestingly, the liposomally encapsulated FITC-S-ODN-PB2–as accumulated in the nuclear region of MDCK cells. However, weak fluorescence was observed within the endosomes and the cytoplasm of MDCK cells treated with the free antisense S-ODNs. The cationic lipid particles may thus be a potentially useful delivery vehicle for oligonucleotide-based therapeutics and transgenes, appropriate for use in vitro or in vivo.
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DOI：
10.1073/pnas.83.17.6282
发表时间：
1986-09-01
影响因子：
11.1
作者：
BEATON, AR;KRUG, RM
通讯作者：
KRUG, RM
DOI：
10.1073/pnas.78.12.7355
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
ULMANEN, I;BRONI, BA;KRUG, RM
通讯作者：
KRUG, RM
与 1 型单纯疱疹病毒立即早期前 mRNA 4 和 5 的剪接点互补的寡核苷酸（甲基膦酸核苷）的抗病毒作用。
DOI：
10.1073/pnas.83.9.2787
发表时间：
1986
影响因子：
11.1
作者：
Smith,CC;Aurelian,L;Reddy,MP;Miller,PS;Ts'o,PO
通讯作者：
Ts'o,PO
DOI：
10.1073/pnas.88.18.8237
发表时间：
1991-09-01
影响因子：
11.1
作者：
PONTIUS, BW;BERG, P
通讯作者：
BERG, P