The article reviews the advancements in CRISPR–Cas technologies and their applications in genome editing and epigenetic modulation. CRISPR–Cas systems, derived from prokaryotes, have revolutionized the manipulation of DNA and RNA sequences in living cells, enabling precise and robust genome editing. The ease of use and versatility of these technologies have expanded their applications from fundamental research to translational medicine. Key advancements include the development of new CRISPR–Cas systems, such as Cas12a and Cascade, which offer broader targeting capacity and unique cutting mechanisms. These systems can be engineered to target specific DNA or RNA sequences, facilitating gene editing and epigenetic modulation. The article discusses the use of CRISPR–Cas tools for gene editing, including non-homologous end joining (NHEJ) and homologous-directed repair (HDR) for DNA repair, as well as single-base editing enzymes. It highlights the development of RNA-targeting tools, such as RCas9 and Cas13, which can be used for RNA recognition and manipulation. These tools have been applied in various contexts, including gene regulation, chromatin reorganization, and therapeutic applications. The review also covers the development of high-throughput screening methods, such as CRISPR-based knockout screens, which have been used to identify essential genes and synthetic lethal gene pairs. These methods have been applied to study cancer and other genetic diseases, providing insights into disease mechanisms and potential therapeutic targets. Finally, the article discusses the biomedical applications of CRISPR–Cas tools, including pre-clinical gene therapy and cell therapy. The potential of CRISPR–Cas technologies in gene therapy is highlighted, emphasizing the ability to make precise, targeted genome modifications for treating genetic disorders.The article reviews the advancements in CRISPR–Cas technologies and their applications in genome editing and epigenetic modulation. CRISPR–Cas systems, derived from prokaryotes, have revolutionized the manipulation of DNA and RNA sequences in living cells, enabling precise and robust genome editing. The ease of use and versatility of these technologies have expanded their applications from fundamental research to translational medicine. Key advancements include the development of new CRISPR–Cas systems, such as Cas12a and Cascade, which offer broader targeting capacity and unique cutting mechanisms. These systems can be engineered to target specific DNA or RNA sequences, facilitating gene editing and epigenetic modulation. The article discusses the use of CRISPR–Cas tools for gene editing, including non-homologous end joining (NHEJ) and homologous-directed repair (HDR) for DNA repair, as well as single-base editing enzymes. It highlights the development of RNA-targeting tools, such as RCas9 and Cas13, which can be used for RNA recognition and manipulation. These tools have been applied in various contexts, including gene regulation, chromatin reorganization, and therapeutic applications. The review also covers the development of high-throughput screening methods, such as CRISPR-based knockout screens, which have been used to identify essential genes and synthetic lethal gene pairs. These methods have been applied to study cancer and other genetic diseases, providing insights into disease mechanisms and potential therapeutic targets. Finally, the article discusses the biomedical applications of CRISPR–Cas tools, including pre-clinical gene therapy and cell therapy. The potential of CRISPR–Cas technologies in gene therapy is highlighted, emphasizing the ability to make precise, targeted genome modifications for treating genetic disorders.