High-throughput Experimental Platforms for Polymeric Vector Discovery

ComboRAFT
image cyt process

Recognizing the urgent clinical need for synthetic vectors, Ramya implemented an unbiased materiomics approach to polymer-mediated gene delivery. She faced two obstacles: 1) the intricacies involved in the rapid synthesis of precisely designed polymers through controlled radical polymerization techniques and 2) the paucity of experimental platforms that unite throughput, analytical rigor and precision.

Ramya approached the tradeoff between analytical depth and sample throughput by engineering a highly scalable workflow wherein a large multiparametric polymer library was rapidly synthesized using a novel platform for parallel RAFT polymerization and then rigorously characterized using automated platforms for evaluating polyplex size, protonation equilibria, and nucleic acid- polymer binding affinities. Finally, these polymers were interrogated through high-content imaging readouts of gene editing efficiency and payload uptake, and image processing pipelines were deployed to identify hit polymers.

Selected Publications

Kumar, R., Le, N., Oviedo, F., Brown, M.E., & Reineke, T.M. (2022) Combinatorial Polycation Synthesis and Causal Machine Learning Reveal Divergent Polymer Design Rules for Effective pDNA and Ribonucleoprotein Delivery. JACS Au, 10.1021/jacsau.1c00467 [PDF]   [Link to Article]
Bockman, M.R., Dalal, R.J., Kumar, R., & Reineke, T.M. (2021) Facile synthesis of GalNAc monomers and block polycations for hepatocyte gene delivery. Polymer Chemistry, 10.1039/D1PY00250C. [PDF]   [Link to Article]
Dalal, R.J., Kumar, R, Ohnsorg, M., Brown, M.E., & Reineke, T.M. (2021) Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression. ACS Macro Letters, 10, XXX, 886–893. [PDF]   [Link to Article]
Kumar, R*., Chalarca, C.F.S.*, Bockman, M.R.*, Van Bruggen, C., Grimme. C.J., Dalal, R.J., Hanson, M.G., Hexum, J.K., & Reineke, T.M. (2021) Polymeric Delivery of Therapeutic Nucleic Acids. Chemical Reviews, 10.1021/acs.chemrev.0c00997. *equal contribution [PDF]   [Link to Article]
Kumar, R., Le, N., Tan, Z., Brown, M.E., Jian, S., & Reineke, T.M. (2020) Efficient polymer-mediated delivery of ribonucleoprotein payloads through combinatorial design & parallelized experimentation. ACS Nano, 10.1021/acsnano.0c08549. [PDF]   [Link to Article]
Tan, Z., Jiang, Y., Ganewatta, M.S., Kumar, R., Keith, A., Twaroski, K., Pengo, T., Tolar, J., Lodge, T.P., Reineke, T.M. Block Polymer Micelles Enable CRISPR/Cas9 Ribonucleoprotein Delivery: Physicochemical Properties Affect Packaging Mechanisms and Gene Editing Efficiency. Macromolecules, 52, 21, 8197-8206 (2019). [PDF]   [Link to Article]