CRISPR/Cas9 Delivery Systems to Enhance Gene Editing Efficiency.

Area of Science:

• Biotechnology
• Molecular Biology
• Gene Therapy

Background:

• CRISPR/Cas9 technology offers precise genome editing with significant therapeutic promise.
• Key challenges include off-target effects, immunogenicity, and efficient delivery of CRISPR components.
• Cas9 protein aggregation can hinder cellular uptake, encapsulation, and nuclear localization, impacting therapeutic efficacy.

Purpose of the Study:

• To provide an integrated analysis of CRISPR/Cas9 delivery systems.
• To highlight advances in physical, viral, and non-viral delivery platforms.
• To identify critical parameters for successful CRISPR/Cas9-based therapies and propose standardized assessment methods.

Main Methods:

• Review of physical, viral, and non-viral delivery systems for CRISPR/Cas9 components.
• Analysis of recent advances in lipid nanoparticles, polymeric carriers, and hybrid platforms.
• Examination of Cas9 protein aggregation and its impact on delivery and editing outcomes.

Main Results:

• Comparison of delivery platforms and their editing outcomes reveals critical physicochemical parameters.
• Identification of Cas9 aggregation as a significant factor affecting delivery efficiency and nuclear localization.
• Insights into challenges related to manufacturing scalability and regulatory requirements for CRISPR/Cas9 therapies.

Conclusions:

• Optimizing delivery systems and addressing Cas9 aggregation are crucial for effective CRISPR/Cas9 therapies.
• Standardized methods for assessing Cas9 encapsulation and aggregation are needed.
• Overcoming translational barriers is essential for the clinical application of CRISPR/Cas9 technology.