Chen Baoling, Pan Ran, Askhatova Diana, Chen P
Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada ; Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON, Canada.
Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada.
Int J Nanomedicine. 2015 May 2;10:3303-14. doi: 10.2147/IJN.S79306. eCollection 2015.
A crucial bottleneck in RNA interference-based gene therapy is the lack of safe and efficient delivery systems. Here, a novel small interfering RNA (siRNA) delivery peptide, STR-HK, was constructed by conjugating a stearyl end to the N-terminus of the peptide sequence HHHPKPKRKV, where PKPKRKV is an altered sequence of the nucleus localization signal (PKKKRKV) and contributes to the cytosol localization of STR-HK-siRNA complexes. Histidine is a linker and plays an important role in disrupting the endosomal membrane via the proton sponge effect. As expected, STR-HK formed complexes with siRNA with a particle size of 80-160 nm in diameter and efficiently delivered Cy3-labeled glyceraldehyde 3-phosphate dehydrogenase siRNA into PC-3 human prostate cancer cells. The transfection efficiency of STR-HK at molar ratio of 60/1 was comparable to that of Lipofectamine 2000, one of the most efficient commercially available transfection reagents. Furthermore, the STR-HK-siRNA complexes exhibited minimal cytotoxicity, which was significantly lower than that of Lipofectamine. Taken together, the strategy of conjugating the stearyl moiety with HHHPKPKRKV as a non-viral siRNA delivery system is advantageous.
基于RNA干扰的基因治疗中的一个关键瓶颈是缺乏安全有效的递送系统。在此，通过将硬脂酰末端连接到肽序列HHHPKPKRKV的N端构建了一种新型小干扰RNA（siRNA）递送肽STR-HK，其中PKPKRKV是核定位信号（PKKKRKV）的改变序列，有助于STR-HK-siRNA复合物的胞质溶胶定位。组氨酸是一个连接子，通过质子海绵效应在内体膜破坏中起重要作用。正如预期的那样，STR-HK与siRNA形成直径为80-160nm的复合物，并有效地将Cy3标记的甘油醛-3-磷酸脱氢酶siRNA递送至PC-3人前列腺癌细胞中。摩尔比为60/1时，STR-HK的转染效率与市售最有效的转染试剂之一Lipofectamine 2000相当。此外，STR-HK-siRNA复合物表现出最小的细胞毒性，显著低于Lipofectamine。综上所述，将硬脂酰部分与HHHPKPKRKV缀合作为非病毒siRNA递送系统的策略具有优势。
