Mechanisms of nanoparticle-mediated siRNA transfection by melittin-derived peptides.
Traditional peptide-mediated siRNA transfection via peptide transduction domains exhibits limited cytoplasmic delivery of siRNA due to endosomal entrapment. This work overcomes these limitations with the use of membrane-destabilizing peptides derived from melittin for the knockdown of NFkB signaling in a model of adult T-cell leukemia/lymphoma. While the mechanism of siRNA delivery into the cytoplasmic compartment by peptide transduction domains has not been well studied, our analysis of melittin derivatives indicates that concurrent nanocomplex disassembly and peptide-mediated endosomolysis are crucial to siRNA transfection. Importantly, in the case of the most active derivative, p5RHH, this process is initiated by acidic pH, indicating that endosomal acidification after macropinocytosis can trigger siRNA release into the cytoplasm. These data provide general principles regarding nanocomplex response to endocytosis, which may guide the development of peptide/siRNA nanocomplex-based transfection.
Computational and Molecular Biophysics, Washington University School of Medicine , St. Louis, Missouri. 63108, United States.
Kirk K
KK
Hou
Hua
H
Pan
Lee
L
Ratner
Paul H
PH
Schlesinger
Samuel A
SA
Wickline
D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D011992:Endosomes / Q000378:metabolism; D008555:Melitten / Q000737:chemistry*; D051379:Mice; D018625:Microscopy, Atomic Force; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D053758:Nanoparticles; D010455:Peptides / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000235:genetics*; D015398:Signal Transduction; D014162:Transfection
D016328:NF-kappa B; D010455:Peptides; D034741:RNA, Small Interfering; D008555:Melitten