Talen technology used to produce a synovial sarcoma cell model

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Ressource 1Télécharger: BIB_BCFD2D120384.P001.pdf (2419.57 [Ko])
Etat: Public
Version: Après imprimatur
Document(s) secondaire(s)
Télécharger: Mémoire no 2497 Annexes Mme El Chammah.pdf (935.55 [Ko])
Etat: Public
Version: de l'auteur⸱e
ID Serval
serval:BIB_BCFD2D120384
Type
Mémoire
Sous-type
(Mémoire de) maîtrise (master)
Collection
Publications
Institution
Titre
Talen technology used to produce a synovial sarcoma cell model
Auteur⸱e⸱s
EL CHAMMAH S.
Directeur⸱rice⸱s
STAMENKOVIC I.
Codirecteur⸱rice⸱s
FUSCO C.
Détails de l'institution
Université de Lausanne, Faculté de biologie et médecine
Statut éditorial
Acceptée
Date de publication
2015
Langue
anglais
Nombre de pages
21
Résumé
Synovial sarcoma (SS) is to this day a devastating malignancy, of which we have very little mechanistic understanding. Two elements characterize SS: the cell of origin, a multipotent myoblast precursor cell, and its signature chromosomal translocation t(X:18) that generates the SYT-SSX fusion gene and corresponding fusion protein. The aim of this project was to produce a TALEN, a tool that could be used to produce SS cells de novo by introducing the translocation into a selected cell line (C3H cells). This may give us insight into the translocation's impact on the cell's physiology early in transformation. The TALEN, composed of a nuclease (Folk I) and an amino acid repeat module array, responsible for the specificity of the nuclease's cut point, was generated using a basic cloning technique. The targeted break point in the murine genome was determined following several criteria including its resemblance to human SS break point and the limitation of off-target effect. Damaged DNA produced by the transfection of C3H cells with the TALEN triggered homologous recombination (HR) thereby introducing the LoxP site into the targeted sequence. The lox P sites were then recognized and recombined by CRE thus producing the translocation between chromosome X and 18. Efficiency and specificity of TALEN cleavage must now be demonstrated by PCR screening. HR's specificity and efficiency, must also be established using T7 Endonuclease I mutation detection assay.
Création de la notice
31/08/2016 15:21
Dernière modification de la notice
20/08/2019 16:31
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