Nucleic acid degradation as barrier to gene delivery: a guide to understand and overcome nuclease activity

Abstract

This review discusses DNases and RNases present in the human body, strategies to reduce the degradation of gene therapeutics after administration and available methods to follow nuclease activity in buffer and living cells.

Integration of Hybridization Chain Reaction and Protein-Binding Amplification for Long-Term Imaging of Intracellular mRNA: Avoiding Signal Fluctuation

“…As we know, there are numerous enzymes coexisting in the cells, , which will affect the stability of nucleic acid probes. Therefore, we analyzed the stability of HPSA-nanobeacon and HCR-nanoprobe in vitro first.…”

“…Until now, numerous spherical nucleic acids that integrate nucleic acid into inorganic or organic nanoparticles have been developed for sensing mRNA, but most of them have failed to be applied intracellularly because of limited sensitivity. To solve this problem, enzyme-free amplification techniques were proposed and the in situ visualization of low abundance mRNA was realized in living cells. − Hybridization chain reaction (HCR) is a typical nonenzymatic signal amplification method and numerous hairpin assemblies based on it have been designed. − However, an inescapable issue is that the chain-like assembly that formed after the reaction would be degraded rapidly by endogenous nuclease, causing the fluctuations in the signal of fluorophore decorated on strands and resulting in a short-lived imaging (Scheme , top). Therefore, there is a difficulty in achieving the long-term fluorescence imaging of mRNA in living cells with high stability.…”

DNA nanotechnology-based nucleic acid delivery systems for bioimaging and disease treatment

“…Although DNA nanostructure-based nucleic acid delivery systems open up new frontiers for biomedical applications, they are still hampered by several recognized challenges. 10,146,147 First of all, nanomaterials based on DNA origami are still difficult to be produced at a large scale for clinical applications, which is primarily due to the heavy price and less yield of DNA origami structures. In consequence, exploring novel cost-efficient strategies for the high-yield synthesis of DNA origami nanomaterials is a critical issue to be addressed in the future.…”

Advances of Layered Double Hydroxide‐Based Materials for Tumor Imaging and Therapy

“…[135] However, during gene therapy, naked gene drugs are susceptible to degradation by nuclease after entering the blood circulation. [136] In addition, some gene drugs such as siRNA itself has a negative charge, due to electrostatic repulsion, it is not easy to penetrate the cell membrane with a negative charge on the surface, and thus there are shortcomings such as large differences between in vivo and out of vivo and poor targeting. [137] In order to solve these problems, it is necessary to find a suitable carrier suitable for transporting gene drugs.…”