CAREER: Multilevel Understanding of Biomaterial-Guided CRISPR Delivery

Non-technical summary

Gene editing tools like CRISPR are transforming biology and medicine. However, ensuring their safety and precision remains a critical challenge. Delivering CRISPR only when and where needed would significantly enhance its effectiveness while minimizing unintended effects.

This project explores how biomaterials can be used to enable precise control over CRISPR's location and timing. Biomaterial-guided CRISPR delivery has the potential to enhance CRISPR’s stability, regulate its release, and target its activity to specific locations, offering several advantages over conventional delivery methods. Despite its potential, significant gaps remain in understanding how the physical and chemical properties of biomaterials influence CRISPR delivery, uptake, and editing efficiency.

This research will address these gaps in knowledge by exploring how tuning the composition of biomaterials and nanoparticles impacts CRISPR delivery. The specific objectives will examine how material properties like charge and porosity control CRISPR release and activity and how cell-material interactions influence CRISPR uptake and function. By systematically studying these factors, this project will provide new insights for designing safer and more effective CRISPR delivery systems, advancing gene editing technology for clinical and research applications.

In addition to its scientific goals, the project integrates education and outreach efforts to increase public understanding of how biomaterials can enhance CRISPR’s safety and effectiveness and engage underrepresented and at-risk students in STEM. By combining cutting-edge research with educational initiatives, this project seeks to drive innovation in gene editing while promoting equity and diversity in science and engineering.

Technical summary

Spatial and temporal control over CRISPR delivery is essential for increasing its efficacy and safety. Harnessing three-dimensional (3D) biomaterials as platforms for material-guided CRISPR delivery is a promising strategy to enhance CRISPR efficacy and minimize off-target effects across a wide range of basic and translational applications. Biomaterial-guided CRISPR delivery offers specific advantages due to enhanced CRISPR cargo protection, defined spatial location, and precise control over temporal action.

Despite these advantages, there is a gap in knowledge regarding how material physicochemical properties and cell-material interactions determine CRISPR delivery rates, cellular uptake, and editing function. This CAREER program aims to understand how the physicochemical properties of nanoparticles and 3D biomaterials, along with material-cell interactions, control CRISPR function.

The central hypothesis is that by tuning the formulation and properties of nanoparticles, the charge and porosity of hydrogels, and cell-material interactions, we can control CRISPR's temporal action, editing efficiency, and off-target effects. The proposed work combines a highly tunable alginate hydrogel strategy with novel peptide-based CRISPR delivery and emerging synthetic biology tools to investigate three specific objectives following a multi-level approach: (1) at the CRISPR delivery level, determine the effect of peptide-based nanoparticle formulation and physiochemical properties on CRISPR ribonucleoprotein (RNP) uptake, activity and functionality; (2) at the material properties level, decouple the individual contributions of material charge and porosity on the temporal action of CRISPR nanoparticles; and (3) at the cell-material interaction level, establish the impact of cell adhesion and proliferation on CRISPR cell uptake and editing efficiency.

Additionally, this CAREER project integrates an educational and outreach program focused on: (1) increasing scientific literacy on how biomaterials can enhance the safety and efficacy of gene editing, and (2) promoting STEM interest and retention among underrepresented and at-risk students.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.