Abstract
The logical AND gate gene circuit based on the CRISPR-Cas9 system can distinguish bladder cancer cells from normal bladder epithelial cells. However, the layered artificial gene circuits have the problems of high complexity, difficulty in accurately predicting the behavior, and excessive redundancy, which cannot be applied to clinical translation. Here, we construct minigene circuits based on the CRISPReader, a technology used to control promoter-less gene expression in a robust manner. The minigene circuits significantly induce robust gene expression output in bladder cancer cells, but have nearly undetectable gene expression in normal bladder epithelial cells. The minigene circuits show a higher capability for cancer identification and intervention when compared with traditional gene circuits, and could be used for in vivo cancer gene therapy using the all-in-one AAV vector. This approach expands the design ideas and concepts of gene circuits in medical synthetic biology.
Highlights
The logical AND gate gene circuit based on the CRISPR-Cas[9] system can distinguish bladder cancer cells from normal bladder epithelial cells
The human uroplakin II gene (hUP II) promoter drove the transcription of the Cas[9] gene, and the human telomerase reverse transcriptase (hTERT) promoter controlled the expression of sgRNA
The traditional gene circuit inhibited the proliferation of normal bladder epithelial cells. These results suggested that the minigene circuit could be used to and strongly inhibit the proliferation of bladder cancer cells
Summary
The logical AND gate gene circuit based on the CRISPR-Cas[9] system can distinguish bladder cancer cells from normal bladder epithelial cells. The minigene circuits show a higher capability for cancer identification and intervention when compared with traditional gene circuits, and could be used for in vivo cancer gene therapy using the all-in-one AAV vector This approach expands the design ideas and concepts of gene circuits in medical synthetic biology. Surgery may not completely remove all tumors, so it is easy to have postoperative recurrence Chemotherapy can damage both cancer and normal cells, resulting in serious side effects in patients. Bladder cancer is an ideal model to test the therapeutic effect of the synthetic gene circuits delivered by gene therapy vectors, and it is possible to provide a biological strategy for treating this disease and efficiently
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