1. DNA Delivery by Virus-Like Nanocarriers in Plant Cells
- Author
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Islam, Reyazul, Youngblood, Marina, Kim, Hye-In, González-Gamboa, Ivonne, Monroy-Borrego, Andrea Gabriela, Caparco, Adam A, Lowry, Gregory V, Steinmetz, Nicole F, and Giraldo, Juan Pablo
- Subjects
Plant Biology ,Biological Sciences ,Medical Biotechnology ,Biomedical and Clinical Sciences ,Gene Therapy ,Biotechnology ,Nanotechnology ,Bioengineering ,Genetics ,Arabidopsis ,Green Fluorescent Proteins ,Gene Transfer Techniques ,Plasmids ,Polyamines ,Protoplasts ,Nanostructures ,DNA ,virus ,nanoparticles ,gene delivery ,protoplasts ,plant genetics ,agriculture ,Nanoscience & Nanotechnology - Abstract
Tobacco mild green mosaic virus (TMGMV)-like nanocarriers were designed for gene delivery to plant cells. High aspect ratio TMGMVs were coated with a polycationic biopolymer, poly(allylamine) hydrochloride (PAH), to generate highly charged nanomaterials (TMGMV-PAH; 56.20 ± 4.7 mV) that efficiently load (1:6 TMGMV:DNA mass ratio) and deliver single-stranded and plasmid DNA to plant cells. The TMGMV-PAH were taken up through energy-independent mechanisms in Arabidopsis protoplasts. TMGMV-PAH delivered a plasmid DNA encoding a green fluorescent protein (GFP) to the protoplast nucleus (70% viability), as evidenced by GFP expression using confocal microscopy and Western blot analysis. TMGMV-PAH were inactivated (iTMGMV-PAH) using UV cross-linking to prevent systemic infection in intact plants. Inactivated iTMGMV-PAH-mediated pDNA delivery and gene expression of GFP in vivo was determined using confocal microscopy and RT-qPCR. Virus-like nanocarrier-mediated gene delivery can act as a facile and biocompatible tool for advancing genetic engineering in plants.
- Published
- 2024