Synthetic Monopartite Peptide That Enables the Nuclear Import of Genes Delivered by the Neurotensin-Polyplex Vector
| Autor: | Daniel Martinez-Fong, Carlos E. Orozco-Barrios, Rasajna Nadella, Rosana Fiorentino-Pérez, Minerva Maldonado-Berny, David Reyes-Corona, Bismark Gatica-Garcia, Francisco E. Lopez-Salas, Moreno Mario Gil, Ubaldo García, Maria L. Escobedo-Sanchez, Maria E. Gutierrez-Castillo, Manuel A. Fernandez-Parrilla |
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| Rok vydání: | 2020 |
| Předmět: |
Male
Patch-Clamp Techniques media_common.quotation_subject Transgene Genetic Vectors Active Transport Cell Nucleus Pharmaceutical Science Importin Gene delivery Green fluorescent protein Cell Line Stereotaxic Techniques chemistry.chemical_compound Mice Drug Discovery Animals Receptors Neurotensin Internalization Neurotensin media_common Cell Nucleus Drug Carriers Chemistry Dopaminergic Neurons Gene Transfer Techniques Transfection Genetic Therapy Fusion protein Peptide Fragments Cell biology Rats Models Animal Molecular Medicine Nanoparticles Single-Cell Analysis Plasmids |
| Zdroj: | Molecular pharmaceutics. 17(12) |
| ISSN: | 1543-8392 |
| Popis: | Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/β pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/β pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy. |
| Databáze: | OpenAIRE |
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