Genome editing methods in animal models

Hyunji Lee,Da Eun Yoon,Kyoungmi Kim

doi:10.1080/19768354.2020.1726462

Hyunji Lee, Da Eun Yoon + Show 1 more

Open Access

https://doi.org/10.1080/19768354.2020.1726462

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Abstract

ABSTRACT Genetically engineered animal models that reproduce human diseases are very important for the pathological study of various conditions. The development of the clustered regularly interspaced short palindromic repeats (CRISPR) system has enabled a faster and cheaper production of animal models compared with traditional gene-targeting methods using embryonic stem cells. Genome editing tools based on the CRISPR-Cas9 system are a breakthrough technology that allows the precise introduction of mutations at the target DNA sequences. In particular, this accelerated the creation of animal models, and greatly contributed to the research that utilized them. In this review, we introduce various strategies based on the CRISPR-Cas9 system for building animal models of human diseases and describe various in vivo delivery methods of CRISPR-Cas9 that are applied to disease models for therapeutic purposes. In addition, we summarize the currently available animal models of human diseases that were generated using the CRISPR-Cas9 system and discuss future directions.

Highlights

The clustered regularly interspaced short palindromic repeats (CRISPR) system, which is derived from the adaptive immune system of prokaryotes, is a breakthrough genome editing technology that can be applied to a variety of researches
The cleaved DNA is repaired by non-homologous end joining (NHEJ) or homology directed repair (HDR) endogenous repair mechanisms, to produce insertion or deletion mutations
We address the development of therapeutic approaches using CRISPR-Cas9 in rat models of human diseases

Summary

Introduction

The clustered regularly interspaced short palindromic repeats (CRISPR) system, which is derived from the adaptive immune system of prokaryotes, is a breakthrough genome editing technology that can be applied to a variety of researches. The third-generation gene editing system Streptococcus pyogenes Cas (SpCas9) can efficiently introduce mutations at desired target positions in the genome in a guide RNA (gRNA)dependent manner (Cong et al 2013). SpCas is widely used in a variety of basic research and clinical applications, including in vivo studies, because it provides highly efficient and characterized PAM sequences. AsCpf and LbCpf have high HDR efficiencies, which allow the insertion or conversion of specific sequences These tools can be useful to generate knockin animal models (Moreno-Mateos et al 2017). Base editors are useful gene editing methods that can directly apply human pathogenic singlenucleotide polymorphisms (SNPs) to animal models and therapeutic studies. Intravenous injection Intravenous injection(Hydrodynamic injection) Intravenous injection Intravenous injection (Hydrodynamic injection) Intravenous injection, Intravaginal injection, Retro-orbital injection Stereitactic injection Intravitreal injection Subretinal injection Intravitreal injection Intravitreal injection Intraacranial injection Subretinal injection Intramuscular injection, Intravenous injection Microinjection

Editing Method

LEE ET AL

Findings

Conclusion