"DNA Barcoded Nanotechnology for High Throughput in vivo Nanoparticle Analysis"
James Dahlman, Ph.D.
Wallace H. Coulter Department of Biomedical Engineering
We use chemical engineering, nanotechnology, genomics, and molecular biology to treat disease.
In vivo multigene editing
Using nanoparticles, Cas9, siRNAs, and miRNAs, we can study how many genes work together to promote disease. How do we do this? By delivering multiple siRNAs, miRNAs, or sgRNAs concurrently in vivo. Why is this important? Many diseases are caused by combinations of genes, not a single gene. For example, we co-delivered two RNAs, each targeting a different cancer pathway, to create a 'targeted combination gene therapy' for cancer. We also designed a five gene therapy for heart disease, and reduced inflammation with Cas9 in vivo.
Designing RNAs and proteins for gene editing
Using molecular biology, we rationally design the genetic drugs we want to deliver. How do we do this? For example, using 'dead' guide RNAs, we can simultaneously turn off gene A and turn on gene B in the same cell using CRISPR-Cas9. Why is this important? Precisely turning certain genes on, and others off, will allow us to study how gene combinations promote disease.
High throughput in vivo screens for targeted nanoparticles
We can make thousands of chemically distinct nanoparticles. Historically, we, and others, have tested if nanoparticles work in cell culture, even though systemic drug delivery is extremely difficult to model in a dish. Now, using high throughput microfluidics and DNA barcoding technology, we can study how hundreds of LNPs work in a single mouse. How do we do this? By rapidly 'barcoding' LNPs, and using DNA deep sequencing to measure where they all go at once. For example, the 'heat map' above shows how well 30 LNPs each target 8 tissues in vivo - we measured this at all once. Why is this important? Nanoparticles behave differently in culture and in the body. Now we can try identify LNPs for clinical use.