Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Department of Physiology and Cell Biology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Be'erSheva, Israel.
Dokl Biol Sci. 2023 Dec;513(Suppl 1):S28-S32. doi: 10.1134/S0012496623700862. Epub 2024 Jan 8.
Gene editing using the CRISPR/Cas9 system provides new opportunities to treat human diseases. Approaches aimed at increasing the efficiency of genome editing are therefore important to develop. To increase the level of editing of the CXCR4 locus, which is a target for gene therapy of HIV infection, the Cas9 protein was modified by introducing additional NLS signals and ribonucleoprotein complexes of Cas9 and guide RNA were stabilized with poly-L-glutamic acid. The approach allowed a 1.8-fold increase in the level of CXCR4 knockout in the CEM/R5 T cell line and a 2-fold increase in the level of knock-in of the HIV-1 fusion peptide inhibitor MT-C34 in primary CD4 T lymphocytes.
利用 CRISPR/Cas9 系统进行基因编辑为治疗人类疾病提供了新的机会。因此，开发旨在提高基因组编辑效率的方法非常重要。为了提高 CXCR4 基因座的编辑水平，该基因座是 HIV 感染基因治疗的靶点，通过引入额外的核定位信号，对 Cas9 蛋白进行了修饰，并使用聚-L-谷氨酸稳定了 Cas9 和向导 RNA 的核糖核蛋白复合物。该方法使 CEM/R5 T 细胞系中 CXCR4 敲除水平提高了 1.8 倍，使原代 CD4 T 淋巴细胞中 HIV-1 融合肽抑制剂 MT-C34 的敲入水平提高了 2 倍。
