The treatment of bladder cancer has recently shown minimal progress. Gene therapy mediated by CRISPR provides a new option for bladder cancer treatment. In this study, we developed a versatile liposome system to deliver the CRISPR-Cas13a gene circuits into bladder cancer cells. After in vitro studies and intravesical perfusion studies in mice, this system showed five advantages: (1) CRISPR-Cas13a, a transcriptional targeting and cleavage tool for gene expression editing, did not affect the stability of the cell genome; (2) the prepared liposome systems were targeted to hVEGFR2, which is always highly expressed in bladder cancer cells; (3) the CRISPR-Cas13a sequence was driven by an artificial tumor specific promoter to achieve further targeting; (4) a near-infrared photosensitizer released using near-infrared light was introduced to control the delivery system; and (5) the plasmids were constructed with three crRNA tandem sequences to achieve multiple targeting and wider therapeutic results. This tumor cell targeting lipid delivery system with near-infrared laser-controlled ability provided a versatile strategy for CRISPR-Cas13a based gene therapy of bladder cancer.

中文翻译：

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基于多功能脂质的CRISPR-Cas13a膀胱癌基因治疗基因传递系统。

膀胱癌的治疗最近显示出最小的进展。CRISPR介导的基因治疗为膀胱癌的治疗提供了新的选择。在这项研究中，我们开发了一种多功能脂质体系统，可将CRISPR-Cas13a基因电路传递到膀胱癌细胞中。经过对小鼠的体外研究和膀胱内灌注研究，该系统显示出五个优点：（1）CRISPR-Cas13a，一种用于基因表达编辑的转录靶向和切割工具，不影响细胞基因组的稳定性；（2）将制备的脂质体系统靶向hVEGFR2，该hVEGFR2在膀胱癌细胞中始终高表达。（3）CRISPR-Cas13a序列由人工肿瘤特异性启动子驱动以实现进一步的靶向；（4）引入使用近红外光释放的近红外光敏剂以控制递送系统；（5）用三个crRNA串联序列构建质粒，以实现多重靶向和更广泛的治疗效果。这种具有近红外激光控制能力的靶向肿瘤细胞脂质输送系统为基于CRISPR-Cas13a的膀胱癌基因治疗提供了一种通用策略。