OWENS LAB
Advancing Genome Editing for Precision and Safety
Our lab is focused on overcoming critical challenges in genome editing to enable precise and efficient genetic modifications for therapeutic and biotechnological applications.
By leveraging the distinct capabilities of integrases and transposases, we aim to address limitations in existing technologies, particularly for the safe and targeted integration of large DNA cargos.
Challenges in Genome Editing
Genome editing methods, including nuclease-based technologies such as CRISPR/Cas, often rely on double-stranded DNA breaks (DSBs). These breaks can result in unintended large deletions, genomic rearrangements, and a selective pressure for cells with impaired p53 function. Additionally, current homology-directed repair (HDR)-based methods are inefficient in non-dividing cells and poorly suited for the integration of large transgene payloads, limiting their utility in many therapeutic and industrial settings.
Our Approach:
Transposases and Integrases
We focus on developing and optimizing both transposases and integrases to expand the genome editing toolkit.
•Transposases
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Transposases, such as piggyBac, enable scarless insertion of genetic material into TTAA sites with high efficiency and large cargo capacity. In our lab, we have engineered piggyBac variants tethered to programmable DNA-binding domains, such as dCas9 and TALEs, to achieve site-specific transposition into genomic safe harbors. By refining the DNA-binding specificity of these systems, we are reducing off-target activity while improving precision, broadening their applicability for gene therapy and functional genomics.
•Integrases
Serine integrases, including PhiC31 and Bxb1, offer a complementary approach for precise DNA integration at predefined attachment (att) sites. Using directed evolution techniques such as our phage-assisted continuous evolution (IntePACE) platform, we have developed hyperactive integrase variants capable of integrating large therapeutic transgenes with significantly improved efficiency and specificity. This approach eliminates the reliance on DSBs, reducing risks of genomic instability and enabling applications in diverse cell types.
Applications and Impact
The tools and technologies developed in our lab aim to address fundamental challenges in genome editing, with applications in:
•Gene therapy, including the delivery of therapeutic genes to non-dividing cells.
•Biomanufacturing, through the creation of stable engineered cell lines for pharmaceutical production.
•Synthetic biology, by enabling precise genome engineering for complex biological systems.
By developing innovative solutions to longstanding technical barriers, we are contributing to safer and more effective genome editing methodologies with broad implications across biotechnology and medicine.
