Ashley Blake

Ashley Blake

Biography

Ashley Blake is a senior at Clemson University pursuing a B.S. in Chemical Engineering
with a concentration in Biomolecular Engineering. This summer, she is an MSRP intern in
the Bathe laboratory, where she is investigating DNA origami as a platform for gene therapy
delivery. Her interest in drug delivery began during the COVID-19 pandemic when she
realized that the success of mRNA vaccines depended as much on the design of the delivery
vehicle as the therapeutic itself. Ashley has conducted research spanning biomaterials, drug
delivery, and gene therapy. She also participated in the DAAD RISE program in Germany and
was named a 2026 Goldwater Scholar. She is particularly interested in developing technologies
that improve the delivery of genetic medicines for neurological diseases. Outside the lab, she
tutors and mentors engineering students, supporting their academic and professional growth.

Using CRISPR-Cas9 to Improve Nuclear Localization of Gene-Encoding
DNA Origami
Ashley Blake1, Grant Knappe2, Anna Romanov2, and Mark Bathe2
1Department of Chemical and Biomolecular Engineering, Clemson University
2Department of Biological Engineering, Massachusetts Institute of Technology

Gene-encoding DNA origami is an emerging platform for non-viral gene delivery
that packages therapeutic genes within programmable DNA nanostructures. However, these
nanostructures must reach the nucleus before the encoded genes can be expressed, a process
that remains particularly challenging in non-dividing cells. Although nuclear localization
represents a major barrier to this technology, strategies to overcome it remain limited. We
therefore investigated whether CRISPR-Cas9 ribonucleoproteins (RNPs), which contain
nuclear localization signals, could enhance nuclear localization of gene-encoding DNA
origami. To test this hypothesis, we engineered gene-encoding DNA origami functionalized
with CRISPR-Cas9 RNPs. The resulting nanostructures were successfully assembled and
characterized by gel electrophoresis, dynamic light scattering, fluorescence spectroscopy,
and transmission electron microscopy, confirming successful incorporation of CRISPR-Cas9
RNPs. RNP-functionalized and unloaded gene-encoding DNA origami were then compared in
proliferating and cell cycle-arrested Neuro-2a cells. Nuclear localization and expression of an
encoded eGFP reporter are being evaluated by fluorescence microscopy. By addressing a key
barrier to nuclear localization, this strategy could broaden the applications of gene-encoding
DNA origami for non-viral gene delivery.