Dermatology Hospital, Southern Medical University, Guangzhou, 510091, China.
Cancer Research Institute, School of Basic Medical Sciences, State Key Laboratory of Organ Failure Research, National Clinical Research Center of Kidney Disease, Key Laboratory of Organ Failure Research (Ministry of Education), Southern Medical University, Guangzhou, 510515, China.
J Nanobiotechnology. 2024 May 24;22(1):284. doi: 10.1186/s12951-024-02554-w.
CRISPR-based gene therapy offers precise targeting and specific editing of disease-related gene sequences, potentially yielding long-lasting treatment effects. However, efficient delivery remains a significant challenge for its widespread application. In this study, we design a novel short peptide-conjugated bioreducible polymer named TSP as a safe and effective delivery vector for the CRISPR system. Our results show that TSP markedly boosts transcriptional activation and genome editing activities of multiple CRISPR systems as confirmed by decomposition-seq and Deep-seq, which is resulted from its capability in facilitating delivery of plasmid DNA by promoting cellular uptake and lysosomal escape. Additionally, TSP further enhances genome editing of CRISPR by delivery of minicircle DNA, a condensed form of regular plasmid DNA. More importantly, TSP significantly improves delivery and genome editing of CRISPR system in vivo. In summary, our study highlights TSP as a promising delivery tool for CRISPR applications in vivo.
基于 CRISPR 的基因治疗提供了针对疾病相关基因序列的精确靶向和特异性编辑，可能产生持久的治疗效果。然而，高效的递送仍然是其广泛应用的一个重大挑战。在这项研究中，我们设计了一种新型的短肽偶联的生物还原聚合物，命名为 TSP，作为 CRISPR 系统的一种安全有效的递送载体。我们的结果表明，TSP 通过促进细胞摄取和溶酶体逃逸来促进质粒 DNA 的递送，从而显著提高了多种 CRISPR 系统的转录激活和基因组编辑活性，这一点得到了分解测序和深度测序的证实。此外，TSP 通过递送迷你环 DNA（一种常规质粒 DNA 的浓缩形式）进一步增强了 CRISPR 的基因组编辑。更重要的是，TSP 显著提高了 CRISPR 系统在体内的递送和基因组编辑效率。总之，我们的研究强调了 TSP 作为一种有前途的体内 CRISPR 应用的递送工具。
