Self-crosslinked and reducible fusogenic peptides for intracellular delivery of siRNA
Self-crosslinked and reducible fusogenic peptides for intracellular delivery of siRNA
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DOI：
10.1002/bip.21032
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发表时间：
2008-10-01
期刊：
影响因子：
2.9
通讯作者：
Park, Tae Gwan
中科院分区：
文献类型：
--
作者：
Mok, Hyejung;Park, Tae Gwan
为了在不影响细胞毒性的前提下，提高siRNA-聚乙二醇偶联物在含血清条件下的转染率，合成了一种新型的自交联可还原多肽，以稳定形成纳米级的复合物。在温和的DMSO氧化条件下，两端带有两个胱烯残基的融合肽Kala通过二硫键进行交联。将可还原的交联型Kala(CL-Kala)与绿色荧光蛋白(GFP)siRNA形成纳米络合物。对比分析了Kala和CL-Kala形成的各种聚电解质络合物的大小和形态。与裸Kala和聚乙烯亚胺(PEI)相比，CL-Kala表现出更低的细胞毒性，并与siRNA形成更稳定、更致密的聚电解质络合物，这可能是因为它的电荷密度增加。用MDA-MB-435细胞定量评估基因沉默的程度。CL-Kala/siRNA复合体的基因沉默效率与细胞毒PEI相当。在含血清的培养液中，由于聚乙二醇膜的屏蔽作用，CL-Kala/siRNA-聚乙二醇偶联物表现出较好的基因抑制作用。基于自交联型融合多肽的制剂可以作为一种生物相容的高效非病毒载体用于siRNA的传递。(C)2008年威利期刊公司。
A novel self-crosslinked and reducible peptide was synthesized for stable formation of nanoscale complexes with an siRNA-PEG conjugate to enhance transfection efficiency in serum containing condition without compromising cytotoxicity. A fusogenic peptide, KALA, with two cystene residues at both terminal ends was crosslinked via disulfide linkages under mild DMSO oxidation condition. The reducible crosslinked KALA (cl-KALA) was used to form nano-complexes with green fluorescent protein (GFP) siRNA. Size and morphology of various polyelectrolyte complexes formulated with KALA and cl-KALA were comparatively analyzed. cl-KALA exhibited more reduced cell cytotoxicity and formed more stable and compact polyelectrolyte complexes with sIRNA, compared with naked KALA and polyethylenimine (PEI), probably because of its increased charge density. The extent of gene silencing was quantitatively evaluated using MDA-MB-435 cells. cl-KALA/siRNA complexes showed comparable gene silencing efficiency with those of cytotoxic PEI. In a serum containing medium, cl-KALA/siRNA-PEG conjugate complexes exhibited superior gene inhibition because of the shielding effect of PEG on the surface. The formulation based on the self-crosslinked fusogenic peptide could be used as a biocompatible and efficient nonviral carrier for siRNA delivery. (c) 2008 Wiley Periodicals, Inc.