Nanocarriers for nucleic acids transfection and biotechnological applications

Abstract

Introduction. This paper focuses on the synthesis, physicochemical characterization, bioactivity evaluation, and innovative applications of new biohybrid nanomaterials. Sepiolite is a nanofibrous natural silicate that represents a very attractive nanocarrier for nucleic acids delivery into cells, among other innovative applications in life sciences and biotechnology. Methods. The physicochemical characterization and the bioactivity evaluation of biohybrids were performed using advanced techniques and procedures, including AFM, TEM, Confocal Microscopy, TLVM, FTIR, Z-potential measurements, UV-vis, FACS, Western blot, RT-qPCR, among others. Results. New biohybrids were obtained, based on the assembly of nucleic acids with sepiolite nanofibers. The mechanisms of cell internalization and response to sepiolite exposure were proposed. We demonstrated that sepiolite is a promising nanocarrier for the non-viral and stable transfer of plasmid DNA into bacteria, mammalian and human cells, with an optimized efficiency. Due to its ability to bind various biomolecules, sepiolite could be used as a nanocarrier for the simultaneous vectorization of diverse biological molecules, including proteins, nucleases and antibodies. In addition, we obtain new protocols for both, strongly improving bacterial transformation efficiency, and to extract plasmids from bacteria, in a rapid, convenient and inexpensive method that doesn’t require competent cell preparation, representing an advantageous alternative to onerous commercial kits. As a conclusion, Because of the low cost of sepiolite, its low toxicity, its natural fluorescence, the ease and convenience of sepiolite/DNA synthesis methods, the possibilities of improving the transfection efficiency and additional enticing future developments, sepiolite represents a very attractive nanoplatform for potential applications in nanomedicine and nanobiotechnology.