A Semiconducting Polymer NanoCRISPR for Near-Infrared Photoactivatable Gene Editing and Cancer Gene Therapy

Area of Science:

• Biotechnology
• Nanotechnology
• Gene Editing

Background:

• Clustered regularly interspaced short palindromic repeat (CRISPR) gene editing faces challenges with efficacy and off-target effects.
• Developing precise and efficient CRISPR delivery systems is crucial for therapeutic applications.

Purpose of the Study:

• To develop a semiconducting polymer (SP)-based nanoCRISPR system for improved CRISPR delivery and efficacy.
• To enable near-infrared (NIR) photoactivatable gene editing for precise cancer therapy.

Main Methods:

• An amphiphilic SP was functionalized with single-stranded DNA (ssDNA) to form an sgRNA/SP-DNA complex (nanoCRISPR).
• The nanoCRISPR system utilizes the photothermal effect of SPs under NIR irradiation to trigger controlled sgRNA release.
• Gene editing efficacy was demonstrated by regulating the expression of green fluorescent protein (GFP) and polo-like kinase 1 (PLK1).

Main Results:

• The nanoCRISPR system effectively delivered sgRNA into cells.
• NIR laser irradiation induced localized heating, controlling sgRNA release and CRISPR activity.
• Precise regulation of GFP and PLK1 gene expression was achieved, demonstrating the potential for precision gene therapy.

Conclusions:

• The SP-based nanoCRISPR system offers a promising approach for enhancing CRISPR gene editing efficacy and safety.
• NIR photoactivation provides a spatiotemporal control mechanism for gene editing, suitable for targeted cancer therapy.