Genomic tagging of endogenous SYT-SSX fusion protein in synovial sarcoma cell line HS-SYII
Details
State: Public
Version: After imprimatur
License: Not specified
Serval ID
serval:BIB_8E0EF1980F87
Type
A Master's thesis.
Publication sub-type
Master (thesis) (master)
Collection
Publications
Institution
Title
Genomic tagging of endogenous SYT-SSX fusion protein in synovial sarcoma cell line HS-SYII
Director(s)
STAMENKOVIC I.
Institution details
Université de Lausanne, Faculté de biologie et médecine
Publication state
Accepted
Issued date
2018
Language
english
Number of pages
23
Abstract
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.
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.
Keywords
Synovial Sarcoma, Genome editing, SYT-SSX, Cell culture, HS-SYII
Create date
03/09/2019 9:23
Last modification date
08/09/2020 7:09
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