Targeted gene integration in skeletal muscle – Muscle represents a critical tissue for gene replacement. Over 70 neuromuscular diseases are cause by genetic mutations in muscle. Designing a versatile gene therapy to address these diseases requires new approaches. We are investigating hijacking of endogenous genes for sustained gene therapy. Moreover, this approach could be used to turn muscle into a protein “factory”.
Relevant work:
Padmaswari, M. H., Bulliard, G., Agrawal, S., Jia, M. S., Khadgi, S., Murach, K. A., & Nelson, C. E. (2024). Precision and efficacy of RNA-guided DNA integration in high-expressing muscle loci. Molecular Therapy Nucleic Acids, 35(4).
Jia MS, Padmaswari MH, Burcham LA, Agrawal S, Bulliard GN, Stokes AL, Nelson CE. Beyond the Cut: Long-read sequencing reveals complex genomic and transcriptomic changes in AAV-CRISPR therapy for Duchenne Muscular Dystrophy. bioRxiv 2025
Delivery of genome editing technologies – As therapeutics, genome editing technologies hold great promise. The challenge is safe and efficient delivery to diseased tissues in the patient. We are adapting viral and non-viral delivery vehicles and assessing safety and efficiency in preclinical models of disease.
Relevant work:
Agrawal S, Stokes AL, Nelson CE. Standardizing a Protocol for Streamlined Synthesis and Characterization of Lipid Nanoparticles to Enable Preclinical Research and Education. bioRxiv 2025
Agrawal S, Padmaswari MH, Stokes AL, Maxenberger DA, Reese M, Khalil A, Nelson CE. “Optimizing Recombinant Cas9 Expression: Insights from E. coli BL21 (DE3) Strains for Enhanced Protein Purification and Genome Editing.” Biomedicines 12, no. 6 (2024): 1226.
Padmaswari MH, Agrawal S, Jia MS, Ivy A, Maxenberger DA, Burcham LA, Nelson CE. Delivery challenges for CRISPR-Cas9 genome editing for Duchenne muscular dystrophy. Invited review for Biophysics reviews. 2023. 4(1), 011307.
Nelson & Gersbach. Engineering Delivery Vehicles for Genome Editing. ARCBE. 2016
Gene regulation in regenerative medicine and wound healing – Changes to gene expression in chronic disease delaying or preventing tissue regeneration or restoration. Targeted control of gene expression can illuminate new drug targets (Review, 2014).
Relevant work:
Ivy A, Bess SN, Agrawal S, Kochar V, Muldoon TJ, and Nelson CE. A Dual-Fluorescence Assay for Gene Delivery Vehicle Screening in Macrophages with an Inflammation-Inducible Reporter Construct. 2025. BMC Methods, 2(1), 1-13
