Protocol for viral vector-mediated measurement of transcription factor activity of mouse brain

Here, we provide a step-by-step protocol to measure the activities of multiple transcription factors (TFs) in the same mouse brain. This protocol includes a procedure to construct a virus-based TF activity reporter, in utero transfection, and PCR-based measurement of TF activity to obtain the transc...

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Veröffentlicht in:STAR protocols 2022-09, Vol.3 (3), p.101633-101633, Article 101633
Hauptverfasser: Yamamoto, Hajime, Abe, Kentaro
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Sprache:eng
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Zusammenfassung:Here, we provide a step-by-step protocol to measure the activities of multiple transcription factors (TFs) in the same mouse brain. This protocol includes a procedure to construct a virus-based TF activity reporter, in utero transfection, and PCR-based measurement of TF activity to obtain the transcription factor activity profile (TFAP). Our protocol facilitates a systematic analysis of TF activity of the brain in vivo and will aid trans-omics understanding of the molecular mechanism underlying the brain functions. For complete details on the use and execution of this protocol, please refer to Abe and Abe, (2022). [Display omitted] •Protocol allows quantitative measurement of TF activity in mouse brain•A protocol for preparation and in utero injection of LV-based transcription reporter•Workflow of behavioral experiment and brain sampling for TF activity measurement Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we provide a step-by-step protocol to measure the activities of multiple transcription factors (TFs) in the same mouse brain. This protocol includes a procedure to construct a virus-based TF activity reporter, in utero transfection, and PCR-based measurement of TF activity to obtain the transcription factor activity profile (TFAP). Our protocol facilitates a systematic analysis of TF activity of the brain in vivo and will aid trans-omics understanding of the molecular mechanism underlying the brain functions.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2022.101633