Generating a GPC-1 Knockout in Glioblastoma Cells Using CRISPR

Start Date

15-4-2021 2:15 PM

End Date

15-4-2021 2:30 PM

Student's Major

Biochemistry

Student's College

Science, Engineering and Technology

Mentor's Name

Samantha Katner

Mentor's Department

Biochemistry

Mentor's College

Science, Engineering and Technology

Description

Glioblastoma (GBM) is a type of aggressive brain cancer that represents roughly 15% of all brain tumors and as of right now, is incurable. GBM is especially known for having abnormal quantities of extracellular matrix proteins called proteoglycans, which are the main protein substituents in brain tissue that serve in development, brain function, and more. A family of proteoglycans, called glypicans (GPCs), play an important role in tumor behavior, and Glypican-1 (GPC-1) has shown to be over produced in glioblastoma cancer cells. Over-expression of GPC-1 causes an increase in a specific growth factor which can overall contribute to tumor growth. Since GPC-1 is over-expressed in GBM cells, generating a knockout (removal) of this protein could then allow us to establish a relationship between GPC-1 and the growth of glioblastoma cancer seen in the clinic. This can be achieved using CRISPR and clonal expansion techniques. CRISPR is a type of gene-editing application that is paired with a Cas-9 endonuclease, which cuts genomic DNA at a specific location, guided by a single guide RNA. This CRISPR technology was introduced to the GL261 cells in order to remove the GPC-1 gene, a clonal expansion was performed to grow up the “pool” of cells transfected with the CRISPR components, and the cells’ DNA was then sent out for sequencing. The GPC-1 gene was successfully knocked out and two true wild-type GBM populations were also obtained for use in further experimentation.

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Apr 15th, 2:15 PM Apr 15th, 2:30 PM

Generating a GPC-1 Knockout in Glioblastoma Cells Using CRISPR

Glioblastoma (GBM) is a type of aggressive brain cancer that represents roughly 15% of all brain tumors and as of right now, is incurable. GBM is especially known for having abnormal quantities of extracellular matrix proteins called proteoglycans, which are the main protein substituents in brain tissue that serve in development, brain function, and more. A family of proteoglycans, called glypicans (GPCs), play an important role in tumor behavior, and Glypican-1 (GPC-1) has shown to be over produced in glioblastoma cancer cells. Over-expression of GPC-1 causes an increase in a specific growth factor which can overall contribute to tumor growth. Since GPC-1 is over-expressed in GBM cells, generating a knockout (removal) of this protein could then allow us to establish a relationship between GPC-1 and the growth of glioblastoma cancer seen in the clinic. This can be achieved using CRISPR and clonal expansion techniques. CRISPR is a type of gene-editing application that is paired with a Cas-9 endonuclease, which cuts genomic DNA at a specific location, guided by a single guide RNA. This CRISPR technology was introduced to the GL261 cells in order to remove the GPC-1 gene, a clonal expansion was performed to grow up the “pool” of cells transfected with the CRISPR components, and the cells’ DNA was then sent out for sequencing. The GPC-1 gene was successfully knocked out and two true wild-type GBM populations were also obtained for use in further experimentation.