Grant: Master's Award: Canada Graduate Scholarships

Genetic knockout is an indispensable tool for studying cellular functions. Recent genomic engineering advancements have led to the gene-editing technology called CRISPR (clustered regularly interspaced short palindromic repeat). This system utilizes a single guided RNAs (sgRNAs) to target a specific gene of interest and effectively induce knockout in nearly all organisms. While CRISPR has been widely applied, it faces limitations such as inconsistent knockout efficiency, effective knockout of only a single gene, and off-target effects. Thus, modification of the CRISPR system is required to solve these issues. The Hippo signaling pathway is a multifaceted gene network that regulates cell proliferation and apoptosis. Signaling is most heavily reliant on phosphorylation and dephosphorylation by kinases and phosphatases respectively, where mutations can lead to tissue overgrowth and cancer. Dr. Yang’s lab has recently identified novel regulators of the Hippo pathway in initiating tumorigenesis, notably PTPN12. We hypothesize that PTPN12 downregulation contributes oncogenic responses in mammalian cells through dysregulation of the Hippo pathway. To elucidate the role of PTPN12, we will develop an inducible Cas9 (iCas9) using Doxycycline (Dox)-inducible system established in Yang lab. Constructs expressing sgRNAs targeting PTPN12 will be made to induce knockout in non-tumorigenic mammary breast cells before quantitative assessment. Enhanced cancer cell proliferation is expected in PTPN12 knockout cells in comparison to wild-type cells. The proposed research will establish the first inducible gene-editing system to study the role of PTPN12 involved in tumorigenesis and will have a significant impact for effective cancer therapy.