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.