Tumor targeted genome editing mediated by a multi-functional gene vector for regulating cell behaviors

Area of Science:

• Biotechnology
• Cancer Research
• Nanomedicine

Background:

• Effective gene delivery is crucial for cancer therapy and prevention.
• Genome editing offers precise control over cellular behavior.
• Developing targeted delivery systems for CRISPR-Cas9 is a key challenge.

Purpose of the Study:

• To construct multifunctional nanoparticles for targeted delivery of CRISPR-Cas9 to tumor cells.
• To evaluate the efficacy of these nanoparticles in gene editing and cancer modulation.
• To assess the impact of genome editing on tumor-associated proteins and the tumor immune microenvironment.

Main Methods:

• Construction of self-assembled nanoparticles using natural polymers (carboxymethyl chitosan).
• Incorporation of CRISPR-Cas9 plasmid, protamine sulfate, endosomolytic peptide (KALA), and AS1411 aptamer.
• In vitro evaluation of cellular uptake, endosomal escape, and gene delivery efficiency.
• Assessment of CDK11 gene knockout and modulation of downstream proteins (MMP-9, VEGF, p53).
• Analysis of immune-related protein expression changes post-genome editing.

Main Results:

• Nanoparticles demonstrated enhanced delivery efficiency and specific targeting of tumor cell nuclei.
• CRISPR-Cas9 mediated by nanoparticles achieved >75% decrease in CDK11 expression.
• Significant down-regulation of MMP-9 and VEGF, and up-regulation of p53 observed.
• Genome editing led to increased expression of immune-related proteins (Fas, CD80, MICA, MICB, HLA-1) and decreased expression of immunosuppressive proteins (CD47, MUC1).

Conclusions:

• Multifunctional nanoparticles effectively deliver CRISPR-Cas9 for targeted gene editing in cancer cells.
• Genome editing via these nanoparticles modulates key proteins involved in tumor development, metastasis, and immune evasion.
• The approach shows promise for reversing tumor-induced immunosuppression and preventing tumor development.