Biological and Biomimetic Material Laboratory (BBML), Center for Sustainable Materials (SusMat), School of Materials Science and Engineering, Nanyang Technological University, 639798, Singapore.
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, China.
ACS Nano. 2023 Sep 12;17(17):16597-16606. doi: 10.1021/acsnano.3c02669. Epub 2023 Aug 16.
CRISPR/Cas9-based genome editing tools have enormous potential for the development of various therapeutic treatments due to their reliability and broad applicability. A central requirement of CRISPR/Cas9 is the efficient intracellular delivery of the editing machinery, which remains a well-recognized challenge, notably to deliver Cas9 in its native protein form. Herein, a phase-separating peptide with intracellular redox-triggered release properties is employed to encapsulate and deliver all three forms of CRISRP-Cas9 editing machinery, namely, pDNA, mRNA/sgRNA, and the ribonucleoprotein complex. These modalities are readily recruited within peptide coacervates during liquid-liquid phase separation by simple mixing and exhibit higher transfection and editing efficiency compared to highly optimized commercially available transfection reagents currently used for genome editing.
基于 CRISPR/Cas9 的基因组编辑工具由于其可靠性和广泛适用性，在各种治疗方法的开发中具有巨大的潜力。CRISPR/Cas9 的一个核心要求是有效地将编辑机制递送到细胞内，这仍然是一个公认的挑战，特别是要以其天然蛋白质形式递 Cas9。在此，采用具有细胞内氧化还原触发释放特性的相分离肽来封装和递呈 CRISPR-Cas9 编辑机制的所有三种形式，即 pDNA、mRNA/sgRNA 和核糖核蛋白复合物。这些形式在液-液相分离过程中通过简单混合很容易被募集到肽凝聚体中，与目前用于基因组编辑的高度优化的商业可用转染试剂相比，具有更高的转染和编辑效率。
