Related Experiment Video
Updated: Nov 29, 2025
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
35.0K
Efficient Polymer-Mediated Delivery of Gene-Editing Ribonucleoprotein Payloads through Combinatorial Design,
Ramya Kumar1, Ngoc Le1, Zhe Tan1
1Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.
ACS Nano
|November 23, 2020
Summary
Researchers developed a novel polymer for efficient delivery of genome-editing tools, surpassing commercial reagents. This breakthrough in polymer design accelerates therapeutic applications for gene editing by identifying key molecular properties for enhanced performance.
Area of Science:
- Biomaterials Science
- Polymer Chemistry
- Gene Editing Technologies
Background:
- Chemically defined polymers offer a promising, non-viral alternative for delivering genome-editing payloads like ribonucleoproteins (RNPs).
- Efficient clinical translation requires rapid exploration of polymer chemical space and understanding structure-function relationships for optimal delivery performance.
Purpose of the Study:
- To discover a polymer enabling efficient intracellular RNP delivery through combinatorial design and high-throughput screening.
- To elucidate structure-function relationships governing polymer performance in RNP delivery, including editing efficiency, cellular toxicity, and RNP uptake.
Main Methods:
- Synthesized a library of 43 statistical copolymers using combinatorial RAFT polymerization, systematically varying monomer pKa, steric bulk, lipophilicity, and hydrophilicity.
- Screened the polymer library using image cytometry to assess RNP delivery and editing efficiency.
- Applied machine learning to analyze structure-function correlations and identify key physicochemical determinants of polymer performance.
Main Results:
- Identified a hit polymer (P38) that achieved nearly 60% nonhomologous end-joining editing efficiency, outperforming commercial transfection reagents.
- Determined that polyplex size distribution and protonation degree influenced cellular toxicity and RNP uptake, respectively.
- Discovered that polymer hydrophobicity and the Hill coefficient (cooperativity-enhanced deprotonation) were critical determinants of editing efficiency, independent of polyplex size and protonation degree.
Conclusions:
- Combinatorial synthesis, high-throughput screening, and data science rapidly identified an optimized polymeric RNP delivery vehicle (P38).
- Established statistically derived design rules highlighting polymer hydrophobicity and deprotonation cooperativity as key factors for enhancing editing efficiency.
- This approach provides a roadmap for designing future polymer libraries tailored for therapeutic RNP-based genome editing applications.
