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Science:刘如谦团队进化出新型基因编辑器EvoCAST,可将整个基因精准高效整合到人类细胞
生物世界· 2025-05-18 01:55
Core Viewpoint - The article discusses advancements in gene editing technology, specifically the development of EvoCAST, a highly efficient and precise system for gene insertion in human cells, which addresses the limitations of existing gene editing methods [2][3][10]. Group 1: Gene Editing Challenges - Integrating entire genes into specific genomic locations has been a long-standing challenge in the field of gene editing [2]. - Existing gene editing technologies can repair most pathogenic gene mutations, but the genetic diversity of many diseases necessitates multiple tailored therapies, limiting patient benefits [2]. Group 2: Discovery of CAST - In June 2019, the discovery of CRISPR-associated transposase (CAST) by teams led by Zhang Feng and Samuel Sternberg marked a significant advancement, allowing for the targeted integration of large DNA segments without causing double-strand breaks [2][3]. Group 3: Development of EvoCAST - The collaboration between Liu Ruqian and Samuel Sternberg led to the evolution of EvoCAST, which significantly enhances the activity of CAST, achieving a 420-fold increase in efficiency for gene insertion in human cells [3][7]. - EvoCAST supports the integration of DNA segments larger than 10kb and can mediate the insertion of therapeutic payloads at various genomic loci related to diseases [3][7]. Group 4: PACE Technology - The PACE (Phage-Assisted Continuous Evolution) technology was utilized to improve the activity of CAST, simulating natural selection to evolve the transposase [5][6]. - After hundreds of rounds of evolution, a variant of the TnsB protein was developed, enhancing integration activity over 200 times without the need for toxic bacterial proteins [6]. Group 5: Comparative Analysis with eePASSIGE - EvoCAST and eePASSIGE, another system developed using PACE, have complementary advantages; eePASSIGE offers higher efficiency, while EvoCAST provides greater editing purity [9][10]. - EvoCAST operates in a single step for gene integration, making it simpler compared to the two-step process required by eePASSIGE [9]. Group 6: Implications for Future Research - The research establishes CAST as a powerful platform for RNA-guided gene integration, suitable for various applications in life sciences and disease treatment [10]. - The study demonstrates how laboratory evolution can transform natural systems into effective therapeutic tools, providing new strategies for improving other CAST systems for efficient gene editing [10].