The most important techniques for gene and genome editing

Document Type : Research Paper

Authors

Department of Agricultural Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

Abstract

Objecrive
This article aims to provide a comprehensive overview of the history of the discovery of these tools, their mechanisms of action, along with their advantages, disadvantages, challenges, applications, and future prospects, presented in a concise and informative manner for enthusiasts.
Materials and methods
Targeted genome editing is based on the induction of double-strand breaks, followed by the repair of the desired strand through mechanisms such as homologous recombination repair (HDR) or non-homologous end joining (NHEJ), which have been utilized across various organisms for diverse purposes. The first specialized nuclease, MegaN, was discovered in 1985, marking the inception of gene editing. Subsequently, the discovery of the ZF motif led to the development of ZFN, TALEN, CRISPR, and Fanzor nucleases, which have been employed for genome editing in living organisms. These nucleases function by binding to the DNA strand, identifying the target sequence, and, through their nuclease domain, introducing a cut. The discovery of the CRISPR/Cas system heralded a new era in gene editing research. The mechanism of action in this system involves the identification of the target sequence by the nuclease through the interaction of DNA and a guide RNA strand; following target sequence identification, the cut is made by the nuclease domain.
Results
Successful examples of these techniques include the treatment of certain diseases: in 2018, Hunter syndrome was treated by introducing the IDS gene into liver cells using the ZFN method in vivo. The TALEN method was used to treat Duchenne muscular dystrophy. In 2021, scientists were able to treat sickle cell anemia using the CRISPR/Cas system through single-base editing techniques.
Conclusions
In modern gene and genome editing methodologies, nuclease proteins are capable of introducing targeted modifications such as insertions, deletions, or substitutions within nucleotide sequences, thereby editing genetic information and the epigenome. These tools are capable of editing pathogenic genes by silencing them or activating inhibitory genes, finding applications in agriculture, medicine, and the treatment of genetic diseases. This system is also effective in gene tagging and modulating gene expression levels. With the development of newer and more precise methods, the use of MegaN nucleases has declined, and research has increasingly focused on TALEN and CRISPR/Cas methods.

Keywords

References

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Agricultural Biotechnology Journal
Volume 18, Issue 2
February 2026
Pages 41-68

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