Versatile Redox-Responsive Polyplexes for the Delivery of Plasmid DNA, Messenger RNA, and CRISPR-Cas9 Genome-Editing Machinery.
Versatile Redox-Responsive Polyplexes for the Delivery of Plasmid DNA, Messenger RNA, and CRISPR-Cas9 Genome-Editing Machinery.
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DOI：
10.1021/acsami.8b09642
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发表时间：
2018-09-26
影响因子：
9.5
通讯作者：
Gong S
中科院分区：
文献类型：
--
作者：
Wang Y;Ma B;Abdeen AA;Chen G;Xie R;Saha K;Gong S
基因治疗为许多疾病的治疗带来了巨大的希望，但由于缺乏安全有效的基因递送系统，基因治疗的临床推广一直很缓慢。在这里，我们报告了两种多功能的氧化还原反应多重体(即，交联型和非交联型)，能够有效地传递各种负电荷有效载荷，包括质粒DNA(DNA)、信使RNA(MRNA)、Cas9/gRNA核糖核蛋白(RNP)和RNP供体DNA复合体(S1mplex)，而没有任何可检测到的细胞毒性。这两类聚合物的关键成分都是一种在主链上含有二硫键并含咪唑基的阳离子聚(N，N‘-双(丙烯酰基)半胺-聚(氨基烷基))聚合物(N，N’-N’-bis(acryloyl)cystamine-co-triethylenetetramine)(p(BAc-Tet)。这种组合物能够实现有效的包裹、细胞摄取、受控的内切/溶酶体逃逸，以及细胞内负电荷有效载荷的解包。为了进一步提高非交联型PBAP聚合物的稳定性，将金刚烷(AD)和β-环糊精(β-CD)偶联到PBAP聚合物上。在多阴离子聚合物如血清白蛋白存在下，β-CD与AD主客体相互作用形成的交联型PBAP复合体比非交联型PBAP复合体更稳定，提示在生理条件下稳定性增强。交联型和非交联型聚合物都显示出相似或更好的转基因和基因组编辑效率，并且生物相容性明显好于具有细胞毒性的商业上可获得的最先进的转染剂Lipofetamine2000。
Gene therapy holds great promise for the treatment of many diseases, but clinical translation of gene therapies has been slowed by the lack of safe and efficient gene delivery systems. Here, we report two versatile redox-responsive polyplexes (i.e., crosslinked and non-crosslinked) capable of efficiently delivering a variety of negatively charged payloads including plasmid DNA (DNA), messenger RNA (mRNA), Cas9/gRNA ribonucleoprotein (RNP), and RNP-donor DNA complexes (S1mplex) without any detectable cytotoxicity. The key component of both types of polyplexes is a cationic poly(N,N’-bis(acryloyl)cystamine-co-triethylenetetramine) (p(BAC-TET)) polymer (a type of poly(N,N’-bis(acryloyl)cystamine-poly(aminoalkyl)) (PBAP) polymer) containing disulfide bonds in the backbone and bearing imidazole groups. This composition enables efficient encapsulation, cellular uptake, controlled endo/lysosomal escape, and cytosolic unpacking of negatively-charged payloads. To further enhance the stability of non-crosslinked PBAP polyplexes, adamantane (AD) and β-cyclodextrin (β-CD) were conjugated to the PBAP-based polymers. The crosslinked PBAP (CLPBAP) polyplexes formed by host–guest interaction between β-CD and AD were more stable than non-crosslinked PBAP polyplexes in the presence of polyanionic polymers such as serum albumin, suggesting enhanced stability in physiological conditions. Both crosslinked and non-crosslinked polyplexes demonstrated either similar or better transfection and genome editing efficiencies, and significantly better biocompatibility than Lipofectamine 2000, a commercially available state-of-the-art transfection agent that exhibits cytotoxicity.
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影响因子：
16.6
作者：
Carlson-Stevermer J;Abdeen AA;Kohlenberg L;Goedland M;Molugu K;Lou M;Saha K
通讯作者：
Saha K
影响因子：
8
作者：
Fröhlich E
通讯作者：
Fröhlich E
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
基于化学方法和纳米载体的基因传递技术研究进展
影响因子：
12.4
作者：
Jin L;Zeng X;Liu M;Deng Y;He N
通讯作者：
He N
影响因子：
10.8
作者：
Moret, I;Peris, JE;Aliño, SF
通讯作者：
Aliño, SF