Abstract: Femtoinjection has been proposed as a feasible approach for the targeted delivery of a decoy oligodeoxynucleotide (ODN) into a single ES cell for the study of transcription factor activity. Here, we evaluated the utility of decoy ODN delivery via femtoinjection in an ES cell model in which Venus fluorescent protein was expressed under… Abstract: Femtoinjection has been proposed as a feasible approach for the targeted delivery of a decoy oligodeoxynucleotide (ODN) into a single ES cell for the study of transcription factor activity. Here, we evaluated the utility of decoy ODN delivery via femtoinjection in an ES cell model in which Venus fluorescent protein was expressed under the control of the tet-off system. Femtoinjection of a control decoy (Con-decoy) and a tetracycline response element decoy (TRE-decoy) into the cytoplasm had no apparent effect on Venus fluorescent protein expression; however, femtoinjection of the TRE-decoy into the nucleus successfully suppressed expression of the Venus fluorescent protein. We therefore conclude that it is feasible to suppress the activity of a transcription factor in a single ES cell by the delivery of a decoy ODN into the nucleus using the femtoinjection technique.Graphical Abstract: Femtoinjection has been proposed as a feasible approach for delivery of a decoy oligodeoxynucleotide (ODN) into a targeted single ES cell for the study of transcription factor activity. The transcription factor activity in each target ES cell was suppressed by the introduction of a large amount of the decoy ODN into the nucleus by femtoinjection. As the transcription factor binds to the introduced decoy ODN, the probability of the transcription factor binding to its target site in the genome was reduced leading to a reduction in activity of the target gene.
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