A Light-Inducible Split-dCas9 System for Inhibiting the Progression of Bladder Cancer Cells by Activating p53 and E-cadherin

Abstract

Optogenetic systems have been increasingly investigated in the field of biomedicine. Previous studies had found the inhibitory effect of the light-inducible genetic circuits on cancer cell growth. In our study, we applied an AND logic gates to the light-inducible genetic circuits to inhibit the cancer cells more specifically. The circuit would only be activated in the presence of both the human telomerase reverse transcriptase (hTERT) and the human uroplakin II (hUPII) promoter. The activated logic gate led to the expression of the p53 or E-cadherin protein, which could inhibit the biological function of tumor cells. In addition, we split the dCas9 protein to reduce the size of the synthetic circuit compared to the full-length dCas9. This light-inducible system provides a potential therapeutic strategy for future bladder cancer.

Programmable Transcriptional Modulation with a Structured RNA-Mediated CRISPR-dCas9 Complex

“…For example, it has been proven that the high expression of P53, E-cadherin, and hBax has a profound effect on bladder cancer cell proliferation, invasion, and apoptosis. 43,44 Therefore, selective activation of those genes responding to upregulated microRNAs in bladder cancer cells may facilitate the development of the bladder cancer therapeutic strategy.…”

“…Moreover, applying the bridge RNA-mediated system in response to microRNA to Cas9/dCas9-based endogenous genetic engineering will facilitate the identification and modification of particular cells at the genetic level in the future. For example, it has been proven that the high expression of P53, E-cadherin, and hBax has a profound effect on bladder cancer cell proliferation, invasion, and apoptosis. , Therefore, selective activation of those genes responding to upregulated microRNAs in bladder cancer cells may facilitate the development of the bladder cancer therapeutic strategy.…”

Modulating gene expression in breast cancer via DNA secondary structure and the CRISPR toolbox

“…In this example, activation of p53 and E-cadherin had a significant effect on reduction of cell proliferation, invasion and apoptosis. While this particular study was conducted in a bladder cancer model, it serves as a good example of what can be achieved using dCas9 ( 213 ).…”

A logic “OR” gate composed of a blue light‐induced CRISPR/dCas9 system and a Tet‐on system activates cancer suppressor genes to inhibit the growth of cutaneous squamous cell carcinoma cells

“…In previous studies, we have established a light‐induced gene expression device based on dCas9 protein and cryptochrome 2 (CRY2)‐cryptochrome‐interacting basic‐helix‐loop‐helix 1 ( CIB1) photosensitive module. [ 7,11–13 ] Under blue light illumination, CRY2 photoreceptor could form a heterodimer with its specific binding CIB1 protein, then dCas9‐CIB1 protein carried CRY2‐activator domain (AD) to activate the expression of target genes. [ 14 ] By controlling the light, the precise control of gene expression is realized.…”

“…(C)Together, the two systems formed a logical "OR" gate, which output in the presence of either blue light or Dox -loop-helix 1 ( CIB1) photosensitive module. [7,[11][12][13] Under blue light illumination, CRY2 photoreceptor could form a heterodimer with its specific binding CIB1 protein, then dCas9-CIB1 protein carried CRY2-activator domain (AD) to activate the expression of target genes. [14] By controlling the light, the precise control of gene expression is realized.…”