Premium
Investigating and Quantitating RNA Binding to Nanoparticles for Potential Use in Gene Therapy
Author(s) -
Pescador Jorge,
Rodriguez Blanca,
Maeder Corina,
Beall Gary,
Lewis Kevin
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.711.20
Subject(s) - rna , oligonucleotide , chemistry , hydrotalcite , nanoparticle , nucleic acid , dna , biophysics , biomolecule , gene delivery , binding selectivity , intercalation (chemistry) , binding site , nanotechnology , gene , biochemistry , biology , genetic enhancement , materials science , catalysis , inorganic chemistry
Nanoparticles are defined as molecules that are 1‐100 nm in size in at least one dimension, which contributes to their large surface area to volume ratio. Hydrotalcite, montmorillonite, and halloysite are nanoparticles that have garnered attention as potential drug delivery carriers in recent years. The low toxicity levels of these nanoparticles as well as their ability to interact with biomolecules, such as proteins and nucleic acids, is a key feature for gene therapy applications using nanotechnology. Previous studies have shown that single‐stranded and double‐stranded oligonucleotides and plasmid DNAs bind well to these nanoparticles. Binding of RNA, however, has not been extensively studied. With RNAi becoming an important potential gene therapy tool, finding suitable drug delivery vehicles for siRNAs is of crucial importance. In our studies, we have evaluated the binding efficiency of different types of RNAs (ssRNA, dsRNA, and complex RNA structures) to different nanoparticles and compared them to DNA binding studies with the same nanoparticles. In initial studies focusing on the layered double hydroxide hydrotalcite, a combination of physical and molecular biological assays was employed to assess binding to DNAs and RNAs with different structures. In addition, competition and displacement studies were performed using anionic salts such as carbonate, sulfate, and phosphate to further study the exchange capacity and the binding affinity of one over the other of these anions with the RNAs of interest. In the future, we also aim to characterize the mode of binding using X‐ray diffraction and transmission electron microscopy.