title: CRISPR/Cas9 mediated knockout of rb1 and rbl1 leads to rapid and penetrant retinoblastoma development in Xenopus tropicalis.
abstract: Retinoblastoma is a pediatric eye tumor in which bi-allelic inactivation of the Retinoblastoma 1 (RB1) gene is the initiating genetic lesion. Although recently curative rates of retinoblastoma have increased, there are at this time no molecular targeted therapies available. This is, in part, due to the lack of highly penetrant and rapid retinoblastoma animal models that facilitate rapid identification of targets that allow therapeutic intervention. Different mouse models are available, all based on genetic deactivation of both Rb1 and Retinoblastoma-like 1 (Rbl1), and each showing different kinetics of retinoblastoma development. Here, we show by CRISPR/Cas9 techniques that similar to the mouse, neither rb1 nor rbl1 single mosaic mutant Xenopus tropicalis develop tumors, whereas rb1/rbl1 double mosaic mutant tadpoles rapidly develop retinoblastoma. Moreover, occasionally presence of pinealoblastoma (trilateral retinoblastoma) was detected. We thus present the first CRISPR/Cas9 mediated cancer model in Xenopus tropicalis and the first genuine genetic non-mammalian retinoblastoma model. The rapid kinetics of our model paves the way for use as a pre-clinical model. Additionally, this retinoblastoma model provides unique possibilities for fast elucidation of novel drug targets by triple multiplex CRISPR/Cas9 gRNA injections (rb1 + rbl1 + modifier gene) in order to address the clinically unmet need of targeted retinoblastoma therapy.
authors.affiliation: Developmental Biology Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: Thomas
authors.initials: T
authors.lastname: Naert
authors.affiliation: Developmental Biology Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: Robin
authors.initials: R
authors.lastname: Colpaert
authors.affiliation: Developmental Biology Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: Tom
authors.initials: T
authors.lastname: Van Nieuwenhuysen
authors.affiliation: Developmental Biology Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.
authors.forename: Dionysia
authors.initials: D
authors.lastname: Dimitrakopoulou
authors.affiliation: Inflammation Research Center, VIB, Ghent, Belgium.
authors.forename: Jannick
authors.initials: J
authors.lastname: Leoen
authors.affiliation: Inflammation Research Center, VIB, Ghent, Belgium.
authors.forename: Jurgen
authors.initials: J
authors.lastname: Haustraete
authors.affiliation: Center for Medical Genetics, Ghent University and Ghent University Hospital, Ghent, Belgium.
authors.forename: Annekatrien
authors.initials: A
authors.lastname: Boel
authors.affiliation: Center for Medical Genetics, Ghent University and Ghent University Hospital, Ghent, Belgium.
authors.forename: Wouter
authors.initials: W
authors.lastname: Steyaert
authors.affiliation: Laboratory for Pharmaceutical Biotechnology, Ghent University, B-9000 Ghent, Belgium.
authors.forename: Trees
authors.initials: T
authors.lastname: Lepez
authors.affiliation: Laboratory for Pharmaceutical Biotechnology, Ghent University, B-9000 Ghent, Belgium.
authors.forename: Dieter
authors.initials: D
authors.lastname: Deforce
authors.affiliation: Center for Medical Genetics, Ghent University and Ghent University Hospital, Ghent, Belgium.
authors.forename: Andy
authors.initials: A
authors.lastname: Willaert
authors.affiliation: Department of Pathology, Ghent University and Ghent University Hospital, Ghent, Belgium.
authors.forename: David
authors.initials: D
authors.lastname: Creytens
authors.affiliation: Developmental Biology Unit, Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium.|Center for Medical Genetics, Ghent University and Ghent University Hospital, Ghent, Belgium.
authors.forename: Kris
authors.initials: K
authors.lastname: Vleminckx
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004195:Disease Models, Animal; D005134:Eye Neoplasms / Q000235:genetics / Q000473:pathology; D055786:Gene Knockout Techniques / Q000379:methods; D012175:Retinoblastoma / Q000235:genetics* / Q000473:pathology; D056286:Retinoblastoma Binding Proteins / Q000235:genetics*; D016160:Retinoblastoma Protein / Q000235:genetics; D050720:Retinoblastoma-Like Protein p107 / Q000235:genetics*; D014981:Xenopus / Q000235:genetics*
chemical_list: D056286:Retinoblastoma Binding Proteins; D016160:Retinoblastoma Protein; D050720:Retinoblastoma-Like Protein p107