New Advances in Using Virus-like Particles and Related Technologies for Eukaryotic Genome Editing Delivery

Abstract

The designer nucleases, including Zinc Finger Nuclease (ZFN), Transcription Activator-Like Effector Nuclease (TALEN), and Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated (CRISPR/Cas), have been widely used for mechanistic studies, animal model generation, and gene therapy development. Clinical trials using designer nucleases to treat genetic diseases or cancers are showing promising results. Despite rapid progress, potential off-targets and host immune responses are challenges to be addressed for in vivo uses, especially in clinical applications. Short-term expression of the designer nucleases is necessary to reduce both risks. Currently, delivery methods enabling transient expression of designer nucleases are being pursued. Among these, virus-like particles as delivery vehicles for short-term designer nuclease expression have received much attention. This review will summarize recent developments in using virus-like particles (VLPs) for safe delivery of gene editing effectors to complement our last review on the same topic. First, we introduce some background information on how VLPs can be used for safe and efficient CRISPR/Cas9 delivery. Then, we summarize recently developed virus-like particles as genome editing vehicles. Finally, we discuss applications and future directions.

Virus-Like Particles as a CRISPR-Cas9 Delivery Tool

“…Despite the limitations and due to the many advantages for biomolecule delivery, VLPs have been effectively utilized for CRISPR-Cas9 genome editing in a handful of studies. A literature review by Lyu and Lu (2022) 37 summarizes all 17 different studies performed to date that utilized VLPs to deliver gene editing tools in vitro or in vivo with cells or mice, respectively, indicating that human trials at this point in time are limited, but will undoubtedly be an important area of focus in the field of genome editing within the near future. To provide some perspective on this growing area of research, here we review several notable studies where researchers have used VLPs to deliver molecular components required for CRISPR-Cas genome editing specifically highlighting recent work where either Nanoblades or Lentivirus-VLPs have been successfully implemented to deliver either RNAs encoding Cas9 and the sgRNA or RNPs containing the Cas9 protein and sgRNA complex.…”

Recent advances in stimuli-responsive polymeric carriers for controllable CRISPR/Cas9 gene editing system delivery

“…6 More importantly, unlike earlier gene editor kits with zinc finger nucleases (ZEN) and transcription activator-like effector nucleases (TALENS), as in the CRISPR/Cas9 gene editing group system, the Cas9 protein is a specific CRISPR-associated protein that designs an sgRNA molecule complementary to the target sequence for precise targeting of every new target gene. [7][8][9][10] It has the advantages of being more simple, versatile and efficient while decreasing the negative aspects of off-target effects as well as lowering the modification costs for nuclease proteins. 11 Thus, given the advantages of precise, efficient and adaptable gene edits on mammalian cells, the genome editing system of CRISPR/Cas9 shows great potential for gene therapy, the construction of disease models, the new discovery of drug targets, and the evaluation of drug success, in addition to its application in cancer and hereditary disease treatment research.…”