Genomic tagging of endogenous SYT-SSX fusion protein in synovial sarcoma cell line HS-SYII

Synovial Sarcoma is a soft tissue malignancy harboring a pathognomonic chromosomal translocation t(X;18)(p11.2;q11.2) producing a chimeric translocation protein called SYT-SSX. It is an aggressive tumor type mainly occurring in children and young adults, with a poor prognosis and no specific therapy. A better understanding of the molecular pathogenesis is needed to develop new therapeutic tools. In recent publications, it has been proven that this translocation protein deeply affects the epigenetic pathways of thecell. To better understand these modifications, a ChIP-Seq analysis can be performed, creating the need for a strong antibody target within the sequence of this chimeric protein. In this work, we induced a double strand break using a CRISPR-Cas9 system and designed a Donor DNA sequence with the aim to use the homology-directed repair mechanism to insert a V5 genomic tag in the HS-SYII cell genome. We then started preliminary work to enable precise co-infection with both the specific CRISPR-Cas9 system and our Donor DNA to induce the V5 tag expression in SYT-SSX