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DLK1 gene editing with CRISPR/Cas9 in cell lines derived from human medullary and differentiated thyroid carcinoma

Grant number: 23/16471-4
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): March 01, 2024
Effective date (End): February 28, 2025
Field of knowledge:Biological Sciences - Genetics - Human and Medical Genetics
Principal Investigator:Janete Maria Cerutti
Grantee:Mariana Rocha Belizario
Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
Associated research grant:21/02752-6 - Multiple Endocrine Neoplasia type 2 (MEN 2) and Medullary Thyroid Carcinoma (TCM): new issues in developmental biology, genetics, immunology, epidemiology, mechanisms of disease and clinical management, AP.TEM

Abstract

Medullary thyroid carcinoma (MTC) is a rare type of thyroid cancer, which can be sporadic or hereditary. The hereditary form is strongly associated with variations in the RET gene (REarranged During Transfection). Besides mutations that promote clinical variability, Copy Number Alterations (CNAs) are related to cancer progression. We have identified that the increase in somatic copies of the DLK1 gene, with a consequent increase in expression, is associated with a larger tumor size. This project aims to perform, through the application of the CRISPR/Cas9 system, the knockout of the DLK1 gene in two MTC-derived cell lines (TT and MZ-CRC-1). As a first step, we will confirm the expression of DLK1 in the MTC cell lines by western blot. Once the expression of the DLK1 gene is confirmed, we will evaluate the number of copies of the DLK1 gene in the cell lines to define the efficiency of the knockout. After this step, we will perform the knockout of the DLK1 gene in the MTC cell lines. Lastly, we will evaluate the efficiency of the DLK1 knockout in the two MTC cell lines. This knockout model in MTC cell lines will allow us to investigate, in future projects, the direct effect of DLK1 gene inactivation on the genesis and/or progression of MTC.

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