The Role of CRISPR-Cas9 in Science and Medicine in the Era of Gene Editing

Abstract

The exact manipulation of DNA sequences made possible by genome engineering, which includes genome and gene editing, transforms genetic research. In contrast to conventional techniques that introduce genetic material into genomes at random, contemporary gene editing technologies like CRISPR-Cas9 enable precise modifications at certain genomic regions. CRISPR-Cas9 was made possible by the development of zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), which replaced earlier methods such as restriction endonucleases. CRISPR-Cas9, which derrived from bacterial immune systems, has become a versatile tool for editing eukaryotic genomes since Jennifer Doudna and Emmanuelle Charpentier developed it in 2012. With the aid of a single-guide RNA (sgRNA), the Cas9 nuclease in CRISPR-Cas9 is able to facilitate targeted genetic alterations through either non-homologous end joining (NHEJ) or homology-directed repair (HDR) mechanisms by causing double-strand breaks at specific DNA sequences. Applications in a number of fields, such as correcting disease-causing mutations in human cells and enhancing disease resistance in crops, have been accelerated by its simplicity and programmability. The development of clinical translation and wider applications is hindered by issues including off-target effects and effective delivery systems, despite its potential. Base editing and prime editing are recent developments that seek to improve accuracy and increase the potential of CRISPR-Cas9 beyond traditional gene editing. This review provides a thorough analysis of the history, mechanisms, applications and challenges of CRISPR-Cas9 technology, highlighting both its transformative impact on genetic research and its potential applications in therapeutic and agriculture.