Parent-of-origin dependent gene-specific knock down in mouse embryos.
In mice hemizygous for the Oct4-GFP transgene, the F1 embryos show parent-of-origin dependent expression of the marker gene. F1 embryos with a maternally derived OG2 allele (OG2(mat)/-) express GFP in the oocyte and during preimplantation development until the blastocyst stage indicating a maternal and embryonic expression pattern. F1-embryos with a paternally inherited OG2 allele (OG2(pat)/-) express GFP from the 4- to 8-cell stage onwards showing only embryonic expression. This allows to study allele specific knock down of GFP expression. RNA interference (RNAi) was highly efficient in embryos with the paternally inherited GFP allele, whereas embryos with the maternally inherited GFP allele showed a delayed and less stringent suppression, indicating that the initial levels of the target transcript and the half life of the protein affect RNAi efficacy. RT-PCR analysis revealed only minimum of GFP mRNA. These results have implications for studies of gene silencing in mammalian embryos.
Iqbal
Khursheed
K
Department of Biotechnology, Institute for Animal Breeding, Mariensee, 31535 Neustadt, Germany.
Kues
Wilfried A
WA
Niemann
Heiner
H
D000483:Alleles; D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009865:Oocytes / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019076:Transgenes
D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins