Class
Article
College
College of Engineering
Department
English Department
Faculty Mentor
Keith Roper
Presentation Type
Poster Presentation
Abstract
Millions of patients die every year from a variety of cancers, and side effects from treatments are marginally better than the disease. T-cell therapy is an emerging treatment method with promising results. The aim of this project is to improve the processes of T-cell therapy by demonstrating the effective transduction of HEK293 cells by delivering CRISPR through a viral vector. Viral backbone plasmids are grown in E.coli and used to transfect HEK293 cells along with CRISPR. CRISPR-containing viral vectors are harvested from the supernatant and used to transduce HEK293 cells. Successful induction is proven by resulting cell GFP expression. By lowering cell density in transfection and transduction, we get improved transduction results. Thermo Fisher Scientific is the company funding and supporting this project, they have a current protocol for cell growth, and transfection. Recent data we gathered from following their protocol provided a lower production of viral particles than expected which has led us to review the protocol in an effort to optimize the process.Our plan is to clone plasmid(s) of interest from E coli, then to recover and confirm plasmid purity. Human embryonic kidney cells will then be transduced followed by recovery to confirm purity of viral-vector encapsulating CRISPR. We will then transfect T lymphocytes with a viral vector and Assay for CRISPR-based transformation related to oncolytic activity.
Location
Logan, UT
Start Date
4-8-2022 12:00 AM
Included in
Delivery of CRISPR in T Cell Therapeutics
Logan, UT
Millions of patients die every year from a variety of cancers, and side effects from treatments are marginally better than the disease. T-cell therapy is an emerging treatment method with promising results. The aim of this project is to improve the processes of T-cell therapy by demonstrating the effective transduction of HEK293 cells by delivering CRISPR through a viral vector. Viral backbone plasmids are grown in E.coli and used to transfect HEK293 cells along with CRISPR. CRISPR-containing viral vectors are harvested from the supernatant and used to transduce HEK293 cells. Successful induction is proven by resulting cell GFP expression. By lowering cell density in transfection and transduction, we get improved transduction results. Thermo Fisher Scientific is the company funding and supporting this project, they have a current protocol for cell growth, and transfection. Recent data we gathered from following their protocol provided a lower production of viral particles than expected which has led us to review the protocol in an effort to optimize the process.Our plan is to clone plasmid(s) of interest from E coli, then to recover and confirm plasmid purity. Human embryonic kidney cells will then be transduced followed by recovery to confirm purity of viral-vector encapsulating CRISPR. We will then transfect T lymphocytes with a viral vector and Assay for CRISPR-based transformation related to oncolytic activity.