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A multifunction lipid-based CRISPR-Cas13a genetic circuit delivery system for bladder cancer gene therapy.

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.