Nanoparticle-Mediated Delivery
for Improved Plant Genetic Engineering
Welcome to the forefront of our research initiative, dedicated to advancing plant genetic engineering through the development of nanomaterials capable of biomolecule delivery to crops. Settled at the intersection of nanotechnology and agriculture, our research aims to establish platforms for the precise delivery of biomolecules, such as proteins and nucleic acids, to plant cells that can be further extended to realize crop genetic engineering technologies. To this end, current projects in our lab focus on carbon nanomaterials and Genetically-encoded Delivery Vehicles (GDVs) as promising biomolecule delivery vehicles. By engineering these tools, we aim to enable efficient and ubiquitous delivery of CRISPR reagents to model organisms and important crops for both transient and stable transformation. Our overarching objective is to create groundbreaking technologies to aid plant synthetic biology in elevating crop productivity and sustainability, ultimately contributing to the global effort for enhanced food security and remediation of climate change.
Genetically-Encoded Delivery Vehicles for DNA-free Plant Gene Editing
Nanoparticle-mediated biomolecule delivery remains at the precipice of plant genetic engineering with the potential to unlock impactful technologies such as biological fertilizers and DNA-free gene editing. Most elusive is the ability to efficiently deliver enzymes and other functional proteins to plant cells, a technology that, in the company of revolutionary gene-editing systems such as CRISPR/Cas, would enable DNA-free gene editing with much reduced off-target activity compared to current methods.
To this end, our lab works at the intersection of protein engineering, nanomaterial science, and plant biology, where we specialize in the engineering of self-assembled protein nanostructures as novel nanocarriers for functional protein cargoes to plant cells. Our current projects focus on a Genetically-encoded Delivery Vehicle (GDV) derived from the assembled nanocapsid of Tobacco Mosaic Virus (TMV). GDVs can be controlled in geometry, surface chemistry, and reactivity by engineering the genes encoding their creation. This allows us to realize controllable bioconjugation of protein cargoes to the nanoparticle surface as well as genetically encoded particle-cargo fusions for downstream delivery of functional proteins to plant cells. Through this project, we are excited to apply GDVs for addressing the ultimate challenge of enabling efficient and accurate DNA-free gene editing through the direct delivery of Cas ribonucleoprotein complexes in lab and field conditions.
Carbon Dot-Mediated Delivery
The current lack of efficient tools for targeted biomolecule delivery and controlled cargo release in plants is a significant obstacle in the study of plant biology and genetic engineering. This challenge has become more pressing due to the increasing importance of addressing food insecurity and the climate crisis. With the need to increase crop yields by 56% to sustain a growing population and the impending threat of a 24% reduction in maize yields by 2030, there is a critical need for improved solutions. Plant genetic engineering faces hurdles due to the rigid plant cell wall, which has a strict size exclusion limit and high turgor pressure, making it challenging to transport bio-macromolecules. Existing delivery methods have limitations in plant range and lack specificity in targeting organelles.
In response, the Demirer lab is developing carbon nanoparticles to enable the controlled release of biocargoes in plant cells. This innovation holds promise for enhancing CRISPR-Cas-mediated genetic engineering in both somatic and stem cells in plants, offering a potential solution to current limitations posed in plant genetic manipulation techniques.
Plant Genetic Engineering via Nanoparticle-Mediated Meristem Transformation
The shoot apical meristem (SAM) is located at the growing tip of plants and serves as a reservoir of stem cells, responsible for generating all aboveground plant organs. This makes it an ideal target for achieving heritable gene editing, without the need for tedious and complex tissue culture or regeneration processes. Edited stem cells within the SAM can ultimately give rise to edited germ cells in the flower, resulting in heritable genetic modifications in subsequent generations. Building upon the established synthesis routes of GDVs and carbon nanomaterials in our lab, we are developing a nanoparticle-based delivery system to efficiently deliver and transiently express CRISPR cargoes in the SAM for heritable and transgene-free plant gene editing.
Related publications
Krasley AT., Li Eugene, Galeana Jesus et al. Carbon Nanomaterial Fluorescent Probes and Their Biological Applications.
Chemical Reviews (2024).
