Photo-Removable CRISPR-Cas12a Systems for Programmable Translational Modulation
- 1Hunan Normal University , , ,
Abstract
Messenger RNA (mRNA) has emerged as a powerful tool for developing new therapeutics and biotechnologies due to its inherent ability to express proteins. However, the lack of precise control over protein expression remains a critical limitation for mRNA-based biomedical applications. Herein, we developed a novel photoactivatable mRNA translational modulation strategy that leverages a photoremovable Cas12a-crRNA ribonucleoprotein (RNP) complex as a programmable translational modulator. This RNP consists of Cas12a and a crRNA engineered with photocleavable (PC) linkers in its spacer region. The resulting binary complex binds specifically to the 5′-untranslated region (5′-UTR) of target mRNAs and inhibits translational initiation. Upon light irradiation, the PC-linkers are cleaved, leading to RNP dissociation and rapid restoration of protein expression. This strategy demonstrated robust phototriggered protein expression both in vitro (firefly luciferase) and in living cells (firefly luciferase, EGFP, and mCherry). Furthermore, the therapeutic potential of this approach was demonstrated in Hep3B cells, where phototriggered translation of tumor suppressor p53 mRNA successfully induced apoptosis. Our study demonstrates an innovative attempt at employing photoremovable RNP to modulate mRNA translation and provides distinct insights for rational exploration of mRNA-based precise therapeutics.
Related
- CRISPR-Cas systems as next-generation antimicrobials: a systemic review of mechanisms, delivery strategies, and translational challenges
- Programmable and selective silencing of oncogenic fusion transcripts by CRISPR-Cas13b via breakpoint sequence recognition
- Engineering gRNA for CRISPR biosensing: chemical, structural, and programmable designs for ultrasensitive diagnostics
- Programmable Genome Sensing with Visual, Fluorescence, and Glucometer Readings: Cell-Free In Vitro Translation Approach Using RNA Switch-CRISPR.
- A circRNA-derived IRES-like element enables programmable translational activation of the endogenous chaperone HSP90
BibTeX
@article{li2026photoremovable,
title = {{Photo-Removable CRISPR-Cas12a Systems for Programmable Translational Modulation}},
author = {Sheng Li and Han Li and Yuling Teng and Erhu Xiong and Ronghua Yang},
journal = {JACS Au},
year = {2026},
month = oct,
doi = {10.1021/jacsau.6c01428},
url = {https://pubs.acs.org/jaaucr/article/doi/10.1021/jacsau.6c01428/5455975/Photo-Removable-CRISPR-Cas12a-Systems-for},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - Photo-Removable CRISPR-Cas12a Systems for Programmable Translational Modulation
AU - Sheng Li
AU - Han Li
AU - Yuling Teng
AU - Erhu Xiong
AU - Ronghua Yang
PY - 2026
DA - 2026/10/22
JO - JACS Au
DO - 10.1021/jacsau.6c01428
UR - https://pubs.acs.org/jaaucr/article/doi/10.1021/jacsau.6c01428/5455975/Photo-Removable-CRISPR-Cas12a-Systems-for
AB - Messenger RNA (mRNA) has emerged as a powerful tool for developing new therapeutics and biotechnologies due to its inherent ability to express proteins. However, the lack of precise control over protein expression remains a critical limitation for mRNA-based biomedical applications. Herein, we developed a novel photoactivatable mRNA translational modulation strategy that leverages a photoremovable Cas12a-crRNA ribonucleoprotein (RNP) complex as a programmable translational modulator. This RNP consists of Cas12a and a crRNA engineered with photocleavable (PC) linkers in its spacer region. The resulting binary complex binds specifically to the 5′-untranslated region (5′-UTR) of target mRNAs and inhibits translational initiation. Upon light irradiation, the PC-linkers are cleaved, leading to RNP dissociation and rapid restoration of protein expression. This strategy demonstrated robust phototriggered protein expression both in vitro (firefly luciferase) and in living cells (firefly luciferase, EGFP, and mCherry). Furthermore, the therapeutic potential of this approach was demonstrated in Hep3B cells, where phototriggered translation of tumor suppressor p53 mRNA successfully induced apoptosis. Our study demonstrates an innovative attempt at employing photoremovable RNP to modulate mRNA translation and provides distinct insights for rational exploration of mRNA-based precise therapeutics.
SN - 2691-3704
ER -