Edição do gene TACR1 em células HEK293T utilizando a técnica CRISPR/Cas9

Abstract

TACR1 encodes the receptor of substance P, which has a neuromodulator role and is associated with inflammatory processes. In the context of neurodegenerative diseases such as osteoarthritis, studying the role of this gene in pathophysiology and nociception may be important to validate it as a molecular target for new therapeutic alternatives. The CRISPR/Cas9 techinique has been widely used to edit diverse cell types, resulting in decreasead, increased or blocked gene activity, or by integration of exogenous sequences into the cell DNA, and can be used to elucidate the role of TACR1 gene in osteoarthritis by its inactivaction. That way, the objective of this work was to performTACR1 gene edition in HEK293t cells by CRISPR/Cas9 techinique. The cloning of sgRNAs in. E.coli bacteria (Stbl3 lineage) was successfull and also the transfection of the cell line with the transformed plasmids.