Forsøksdyr: Generation and Characterization of Alkbh5 and Fto knockout mice


Godkjenningsdato 30.07.2018

FORSØKETS FORMÅL
We have generated and characterized Alkbh5 KO mouse model (See project ID #: 3050), and different types of Fto KO/inactivated mouse model have been developed by other groups, for instance by Fischer and colleague [(Nature 458, 894-898(2009)].

FORVENTET SKADEVIRKNING PÅ DYRENE
As single KO, neither Alkbh5 nor Fto KO mice showed any obvious phenotype except for: (1) Alkbh5 KO mice displayed reduced testis size and impaired fertility, and (2) Fto KO mice are lean (have reduced body size compared with their wild-type litter-mates) and homozygous KO mice are infertile. All experiments (DNA, RNA and protein analysis) will be performed after the animals are terminated.

FORVENTET VITENSKAPELIG/SAMFUNNSMESSIG NYTTEVERDI
Since both Fto and Alkbh5 proteins have been shown to catalyze the demethylation of N6-methyladenosine in nuclear RNA, we suspect that the absence of severe phenotypes in either of the single knockout mice could be because of some sort of functional redundancy between the two proteins. Hence, we have generated Alkbh5/Fto heterozygous double KO mice (Project ID 4775) with the aim of studying the functional consequences of the simultaneous loss of both genes. Interestingly, the proportion of homozygous double KO mice derived from these double HE mice breeding is dramatically reduced, indicating a role for these genes in embryonic and/or early neonatal survival. Therefore, we would like to further characterize the Alkbh5/Fto double KO mice model for a better understanding of the in vivo functions of these genes.

HVOR MANGE/HVA SLAGS DYR
Vi har beregnet et behov for 904 mus over en 4-årsperiode (se beregning lenger nede i søknaden)

ERSTATNING/REDUKSJON OG FORBEDRING
Vi har redusert antall dyr til et minimum. Forsøk som utføres for vevsspesifikke endringer, embryoutvikling og fertilitet er ikke mulig å utføre i cellelinjer. Se forøvrig beskrivelse annet sted i søknaden.