Magnetic nanoparticle mediated-gene delivery for simpler and more effective transformation of Pichia pastoris

• Published in: Nanoscale advances Vol. 3; no. 15; pp. 4482 - 4491
• Main Authors: Yildiz, Seyda, Solak, Kubra, Acar, Melek, Mavi, Ahmet, Unver, Yagmur
• Format: Journal Article
• Language: English
• Published: RSC 27.07.2021
• Subjects: Chemistry
• ISSN: 2516-0230; 2516-0230
• DOI: 10.1039/D1NA00079A

The introduction of exogenous DNA into a cell can be used to produce large quantities of protein. Here, we describe a novel gene delivery method for Pichia pastoris based on recombinant DNA delivery using magnetic nanoparticles (MNPs) under magnetic forces. For this purpose, a linear plasmid (pGKB-GFP) containing the Green Fluorescent Protein (GFP) gene is loaded on polyethyleneimine-coated iron oxide (Fe 3 O 4 @PEI) MNPs at doses that are non-toxic to the yeast cells. The pGKB-GFP loaded MNPs combined with enhancer PEI (Fe 3 O 4 @PEI + pGKB-GFP + PEI) are directly transferred to non-competent cells. An effective GFP expression was observed by the selection of antibiotic-resistant yeast cells and heterologous gene integration into the P. pastoris genome was provided. This method, which is very simple, effective, and advanced equipment-free compared to traditional methods, uses smaller amounts of DNA and the process can be performed in a shorter time. The suggested method might also be adapted for the transformation of other yeast species.