Codelivery of DNA and siRNA via arginine-rich PEI-based polyplexes.
Codelivery of DNA and siRNA via arginine-rich PEI-based polyplexes.
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DOI：
10.1021/mp5006883
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发表时间：
2015-02-02
影响因子：
4.9
通讯作者：
Labhasetwar V
中科院分区：
文献类型：
--
作者：
Lu S;Morris VB;Labhasetwar V
在本研究中，我们构建了不同组成的多聚体，用于DNA和小干扰RNA(SiRNA)的联合传递。由于DNA和siRNA具有不同和互补的形态特征(DNA长而卷曲，小干扰RNA短而刚性)，我们假设它们通过复合载体共同传递会促进彼此的转染率。为了验证这一假设，以阳离子聚合物支化聚乙烯亚胺(BPEI)作为标准的转染剂，以本实验室合成的聚乙二醇改性的富含精氨酸的寡肽接枝的bPEI(P(SiDAAr)5P3)为载体进行转染。将不同核酸/聚合物重量比的多聚体分别转染乳腺癌敏感细胞系(MCF-7)和耐药细胞系(MCF-7/ADR)。在MDA-MB-231-Luc-D3H2LN细胞系中检测多个复合体的基因沉默效应。结果表明，与bPEI形成的聚合物相比，P(SiDAAr)5P3形成的聚合物具有更低的毒性。此外，与各自的单核苷酸传递相比，联合传递导致20倍的DNA转染率和2倍的siRNA转染率。耐药细胞的∼转染率是敏感细胞的100倍。共聚焦成像和流式细胞仪数据表明，增强的转染率并不完全依赖于细胞对DNA的摄取，这表明其他机制有助于提高转染率。DNA-co-siRNA传递可能是一种很有前途的治疗方法，可以实现协同效应，因为它可以同时靶向和干扰细胞中的多个调控水平，以阻止和逆转疾病的进展。
In this study, we formulated polyplexes with different compositions for co-delivery of DNA and small-interfering RNA (siRNA). Since DNA and siRNA have distinctive and complementary morphological characteristics (DNA is long and winding and siRNA is short and rigid), we hypothesized that their co-delivery using polyplex would enhance each other's transfection. To test this hypothesis, cationic polymer branched polyethylenimine (bPEI) as a standard transfecting agent and its derivative arginine-rich oligopeptide-grafted bPEI modified with polyethylene glycol (P(SiDAAr)5P3), synthesized in our laboratory, were used as carriers for transfection. Polyplexes at different nucleic acid to polymer weight ratios were characterized for transfection in breast cancer sensitive (MCF-7) and resistant (MCF-7/Adr) cell lines. Gene silencing effect of polyplexes was determined in MDA-MB-231-luc-D3H2LN cell line. The results demonstrated that the polyplexes formed with derivative P(SiDAAr)5P3 show significantly lower toxicity compared to polyplexes formed using bPEI. Further, co-delivery resulted in 20-fold higher DNA transfection and 2-fold higher siRNA transfection as compared to the respective single nucleotide delivery. DNA transfection was ∼100 fold lower in resistant MCF-7/Adr cells than in sensitive MCF-7 cells. Confocal imaging and flow cytometry data demonstrated that enhanced transfection does not solely depend on DNA's cellular uptake, suggesting that other mechanisms contribute to increased transfection. DNA-co-siRNA delivery could be a promising therapeutic approach to achieve synergistic effects, because it can simultaneously target and interfere with multiple regulatory levels in a cell to halt and reverse disease progression.
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影响因子：
14.9
作者：
Read ML;Singh S;Ahmed Z;Stevenson M;Briggs SS;Oupicky D;Barrett LB;Spice R;Kendall M;Berry M;Preece JA;Logan A;Seymour LW
通讯作者：
Seymour LW
影响因子：
17.1
作者：
Rosenholm, Jessica M.;Meinander, Annika;Linden, Mika
通讯作者：
Linden, Mika
影响因子：
4.8
作者：
Ruponen, M;Rönkkö, S;Urtti, A
通讯作者：
Urtti, A
DOI：
10.1016/j.nano.2011.06.022
发表时间：
2012-04-01
影响因子：
5.4
作者：
Tripathi, Sushil K.;Goyal, Ritu;Gupta, Kailash C.
通讯作者：
Gupta, Kailash C.
影响因子：
3.7
作者：
Fischer, D;Bieber, T;Kissel, T
通讯作者：
Kissel, T