Unit:
Κατεύθυνση Αναπαραγωγική-Αναγεννητική ΙατρικήLibrary of the School of Health Sciences
Author:
Tourkodimitri Magdalini-Ioanna
Supervisors info:
Δρακάκης Πέτρος, Καθηγητής, Ιατρική Σχολή, ΕΚΠΑ, Επιβλέπων
Μαυρογιάννη Δέσποινα, Μέλος ΕΔΙΠ, Ιατρική Σχολή, ΕΚΠΑ
Παπαναγιώτου Κωσταντίνος, Ερευνητής Δ΄ βαθμίδας, ΙΙΒΕΑΑ
Original Title:
Μελέτη της ενεργότητας του ενισχυτή AR1 του καρδιακού δείκτη Nkx2.5 σε βλαστικά κύτταρα μυός
Translated title:
Study of the activity of the cardiac marker Nkx2.5’s enhancer, AR1, in mouse Stem Cells
Summary:
A large part of the scientific community, trying to solve the problem of organ inadequacy for transplantations, has managed to create in vivo various tissues and organs, using the technique of blastocyst complementation. The research team of K. Papanagiotou tries to generate hearts, an organ essential for sustaining life, needed by many patients worldwide and one that has not been successfully synthesized yet, using a different genetic approach. This approach does not make use of the blastocyst complementation principle. Instead it relies on the generation of chimeras made up of two genetically modified ES cell line that will segregate in the developing embryos into heart and non-heart tissues. To create these two cell lines, two DNA sequences will be performed in the laboratory, which will be introduced using the CRISPR method at the Hip11 genetic site of the mESC genome (donor-sequence and recipient-sequence). Nkx2.5 is the first known indicator of cardiac genealogy, while AR1 is the first regulatory element of the Nkx2.5 gene that is activated in cardiogenic mesoderm at day E7.5. Both sequences that will be created will contain the Nkx2.5 enhancer, AR1. It is very important for the experimental procedure that the AR1 enhancer is not active in the mESCs earlier than normal, i.e. on day E7.5, as premature expression will trigger a series of actions that will lead to the death of the cells that will receive the recipient sequence. The purpose of this dissertation is to construct the control plasmids, i.e. the plasmids that will be used to study the activity of the AR1 cardiac enhancer of the Nkx2.5 gene.
Main subject category:
Science
Keywords:
Regenerative medicine, Stem cells, Organ transplantation, Blastocyst complementation, Heart, AR1 enchancer, Nkx2.5 gene, CRISPR, Hip11
Number of references:
123
