西湖大学开发相分离递送技术，重塑多种原代细胞CRISPR基因编辑治疗新格局

Core Viewpoint - CRISPR-Cas9 technology, known as "molecular scissors," is a revolutionary tool in biomedical research, offering new strategies for treating genetic diseases and tumors through precise gene editing [1][5]. Group 1: CRISPR-Cas9 Mechanism - The CRISPR-Cas9 system is derived from bacterial immune mechanisms and has been adapted into a powerful gene editing tool for various applications, including gene knockout, insertion, and regulation [5][8]. - Key components include the Cas9 protein, which cuts DNA, and sgRNA, which guides Cas9 to specific gene sequences [7][8]. Group 2: Challenges in Delivery - Despite its powerful editing capabilities, the application of CRISPR-Cas9 in primary immune cells and stem cells faces significant challenges due to the cells' sensitivity and limited in vitro expansion [2][3]. - Current delivery methods, such as chemical transfection and electroporation, have limitations in efficiency and can cause cell damage, while viral vectors pose risks of insertion mutations and immune responses [2][3]. Group 3: ProteanFect Delivery System - The ProteanFect CRISPRMax Ultra transfection kit, developed by West Lake Aggregates and West Lake University, addresses the delivery bottleneck for difficult-to-transfect primary cells by utilizing innovative biomolecular aggregation technology [9][13]. - This system allows for high transfection efficiency and cell compatibility without relying on harsh physical or chemical methods, thus maintaining cell viability and functionality [9][13]. Group 4: Successful Applications - The ProteanFect CRISPRMax Ultra kit has demonstrated high editing efficiency in various primary cells, including mouse and human T cells, achieving mutation rates of 84.3% and 80.1% respectively [14][16]. - Additionally, it has been successfully used to deliver gene editing systems to induced pluripotent stem cells (iPSCs), achieving a base substitution efficiency of 55% [17]. Group 5: Future of Gene Editing - The demand for efficient gene editing in various functional cells is growing, driven by advancements in precision medicine and cell therapy [19][20]. - Innovations like ProteanFect represent a paradigm shift in research, enabling more gentle and efficient interactions with cells while preserving their functionality [19][20].