ESR4: Functional assessment of a cis-regulatory element of ABCA4 in a frog model
Partner
Ghent University, Belgium (www.ugent.be/en; www.debaerelab.com) Supervisor Prof. Dr. K. Vleminckx |
Munevver Burcu Cicekdal
My name is Munevver Burcu Cicekdal and I was the part of the incredible StarT project at UGent, which was a journey that not only deepened my scientific knowledge but also enriched my personal growth. Raised in the vibrant city of Istanbul, I was constantly surrounded by a mosaic of cultures and perspectives, a diversity mirrored in the StarT consortium. This unique blend of individuals from various countries created an enriching environment, much like my own upbringing. After completing my undergraduate studies of Molecular Biology and Genetics at Istanbul University, I achieved my Master’s degree in Biotechnology and dedicated four years as a Research Assistant at Yeditepe University. It was a period filled with learning and discovery, laying a solid foundation for my future endeavours. Joining the StarT project at UGent was a pivotal moment in my career, placing me at the heart of an ambitious effort to tackle Stargardt’s disease. This initiative brought together experts from various fields, all united in their quest to make a breakthrough. As a part of this team, in my capacity as ESR4, I was privileged to work with cutting-edge technology, guided by some of the field's leading minds. My research focused on the complex area of cis-regulatory elements of inherited retinal disease genes in Xenopus tropicalis. This work not only pushed the boundaries of genetic research but also allowed me to contribute to our collective understanding of a challenging genetic disorder. Prospected date to obtain my PhD degree in Biochemistry and Biotechnology: September 2024. Being part of the Marie Skłodowska-Curie Actions Innovative Training Network (MSCA ITN) was the highlight of my scientific journey so far. It was truly enriching to participate in various trainings within an international setting. The secondments were a fantastic opportunity to engage myself in diverse scientific cultures and practices, significantly broadening my perspective. StarT brought together an amazing group of ESRs, and being part of this community was both inspiring and motivating. The experience was not just about scientific discovery but also about personal growth, learning from others, and being part of a collective effort to make a difference. I am excited about what the future holds and am committed to continuing my contribution to the scientific community as a Post-doctoral researcher. |
Abstract
Xenopus tropicalis contains the major cell types of the human eye and unlike the rod-dominated rodent retina, Xenopus (X.) has equal numbers of cones and rods, which is more similar to humans. In addition, abca4 is not duplicated in X. and is located in a region with high synteny, facilitating the identification and characterisation of possible CREs. Previous CRISPR/Cas9 based knock out a CRE of Shh (i.e. ZRS) in X. tropicalis resulted in a phenocopy of the human limb phenotype. This work demonstrated that X. can be used as a model organism to study the non-coding genome. To generate a stable knockout of a previously characterised CRE of ABCA4, ESR4 will use CRISPR/Cas9-mediated genome editing to disrupt this element in X. ESR4 will screen F0 or F1 animals for a STGD1 phenotype by histology, immunofluorescence and TUNEL assays. To study the effect of variations in CREs in F0 X. and to add to the stable transgenesis experiments of ESR2, ESR4 will use a recently described direct plasmid-injection method generating robust, promoter-typical expression in tadpoles. A reference gene present on the plasmid controls for variations in the injections. The plasmids further contain phiC31 AttB sites to favour integration and chromosomal insulators to reduce possible chromosomal position effects.
Xenopus tropicalis contains the major cell types of the human eye and unlike the rod-dominated rodent retina, Xenopus (X.) has equal numbers of cones and rods, which is more similar to humans. In addition, abca4 is not duplicated in X. and is located in a region with high synteny, facilitating the identification and characterisation of possible CREs. Previous CRISPR/Cas9 based knock out a CRE of Shh (i.e. ZRS) in X. tropicalis resulted in a phenocopy of the human limb phenotype. This work demonstrated that X. can be used as a model organism to study the non-coding genome. To generate a stable knockout of a previously characterised CRE of ABCA4, ESR4 will use CRISPR/Cas9-mediated genome editing to disrupt this element in X. ESR4 will screen F0 or F1 animals for a STGD1 phenotype by histology, immunofluorescence and TUNEL assays. To study the effect of variations in CREs in F0 X. and to add to the stable transgenesis experiments of ESR2, ESR4 will use a recently described direct plasmid-injection method generating robust, promoter-typical expression in tadpoles. A reference gene present on the plasmid controls for variations in the injections. The plasmids further contain phiC31 AttB sites to favour integration and chromosomal insulators to reduce possible chromosomal position effects.
