Strategic engineering of DNA aptamers as precision modulators in CRISPR-driven oncogene control

Area of Science:

• Molecular Biology
• Gene Editing Technologies
• Cancer Therapeutics

Background:

• CRISPR-based signal conductor systems are limited by the availability of intracellular protein-binding aptamers.
• Precise and dynamic gene regulation is crucial for effective cancer therapy.

Purpose of the Study:

• To develop a programmable CRISPR-SaCas9 platform using DNA aptamers.
• To achieve precise and dynamic gene regulation for cancer therapy.

Main Methods:

• Identified and characterized five DNA aptamers specific for the SaCas9 protein.
• Engineered aptamers for transcription factor-responsive gene expression control.
• Evaluated therapeutic efficacy in vivo, monitoring tumor progression and metastasis.

Main Results:

• Developed aptamer-CRISPR complexes forming ternary assemblies with SaCas9 and sgRNAs.
• Demonstrated logic-gated gene expression control responsive to endogenous transcription factors.
• Achieved significant suppression of tumor growth and metastasis in vivo.

Conclusions:

• A modular CRISPR signal conductor platform was successfully developed.
• The platform utilizes SaCas9-binding DNA aptamers for transcription factor-mediated logic operations.
• This system offers a promising strategy for targeted cancer gene therapy and expands genetic circuit design tools.