Design and validation of a reporter mouse to study the dynamic regulation of TFEB and TFE3 activity through < em > in vivo < /em > imaging techniques

Autophagy. 2024 Mar 24. doi: 10.1080/15548627.2024.2334111. Online ahead of print.ABSTRACTTFEB and TFE3 belong to the MiT/TFE family of transcription factors that bind identical DNA responsive elements in the regulatory regions of target genes. They are involved in regulating lysosomal biogenesis, function, exocytosis, autophagy, and lipid catabolism. Precise control of TFEB and TFE3 activity is crucial for processes such as senescence, stress response, energy metabolism, and cellular catabolism. Dysregulation of these factors is implicated in various diseases, thus researchers have explored pharmacological approaches to modulate MiT/TFE activity, considering these transcription factors as potential therapeutic targets. However, the physiological complexity of their functions and the lack of suitable in vivo tools have limited the development of selective MiT/TFE modulating agents. Here, we have created a reporter-based biosensor, named CLEARoptimized, facilitating the pharmacological profiling of TFEB- and TFE3-mediated transcription. This innovative tool enables the measurement of TFEB and TFE3 activity in living cells and mice through imaging and biochemical techniques. CLEARoptimized consists of a promoter with six coordinated lysosomal expression and regulation motifs identified through an in-depth bioinformatic analysis of the promoters of 128 TFEB-target genes. The biosensor drives the expression of luciferase and tdTomato reporter genes, allowing the quantification of...

Source: Autophagy - March 24, 2024 Category: Cytology Authors: Electra Brunialti Nicoletta Rizzi Rita Pinto-Costa Alessandro Villa Alessia Panzeri Clara Meda Monica Rebecchi Donato A Di Monte Paolo Ciana Source Type: research