Mentor Areas
Molecular biology, genome engineering, hematopoiesis, pluripotent stem cells.
Description:
Integrating large DNA fragments at specific genomic locations in induced pluripotent stem cells (iPSCs) remains challenging. Serine Site-specific recombinases (SSRs) like Bxb1 are used in synthetic biology to integrate, invert and excise large DNA fragments by promoting the recombination of attP and attB sites. This reaction is irreversible and unidirectional, making this ideal for stably integrating expression cassettes (containing an attB site) into the genome where the landing pad (attP) is present.
Generating a wildtype iPSC line with an attP landing pad would be extremely useful for the field, as it will allow the targeted integration of large DNA payloads and the rapid generation of monoallelic or biallelic transgenic lines, without the introduction of DNA double-stranded breaks.
In this project, we plan to use prime editors to install a landing pad into the B2M locus in an iPSC line. This will allow the precise and efficient Bxb1-mediated insertion of large DNA payloads, including reporter genes, resistance cassettes, and gene variants. This system will boost our ability to generate iPSC-derived disease models and reporter lines in a shorter time, but also provide a platform for high-throughput applications, such as cell libraries and screenings.
Preferred Qualifications
Some experience with cell culture and molecular biology is preferred, but not required.
Details:
Preferred Student Year
First-year, Second-Year, Junior, Senior
Academic Term
Fall, Spring, Summer
I prefer to have students start during the above term(s).Volunteer
Yes
Yes indicates that faculty are open to volunteers.Paid
No
Yes indicates that faculty are open to paying students they engage in their research, regardless of their work-study eligibility.Work Study
Yes
Yes indicates that faculty are open to hiring work-study-eligible students.