Synthesizing AND gate minigene circuits based on CRISPReader for identification of bladder cancer cells

• Published in: Nature communications 2020-10, Vol.11 (1), p.5486-5486, Article 5486
• Main Authors: Liu, Yuchen, Huang, Weiren, Cai, Zhiming
• Format: Article
• Language: English
• Subjects: 13/109; 13/2; 13/44; 38/77; 38/88; 38/90; 42/41; 59/5; 631/1647/1513/1967/3196; 631/61/338/552; 631/67; 64/60; Biology; Bladder; Bladder cancer; Cancer; Circuit design; Circuits; CRISPR; Epithelial cells; Gates (circuits); Gene expression; Gene therapy; Humanities and Social Sciences; Logic circuits; multidisciplinary; Redundancy; Robustness; Science; Science (multidisciplinary); Synthetic biology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-19314-7

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. Synthetic biology logic gates can be used to distinguish healthy cells from cancer cells. Here the authors design minigene circuits that show more robust identification of cancer cells compared to traditional genetic circuits.