Abstract
YAP protein is a critical regulator of mammalian embryonic development. By generating a near-infrared fusion YAP reporter mouse line, we have achieved high-resolution live imaging of YAP localization during mouse embryonic development. We have validated the reporter by demonstrating its predicted responses to blocking LATS kinase activity or blocking cell polarity. By time lapse imaging preimplantation embryos, we revealed a mitotic reset behaviour of YAP nuclear localization. We also demonstrated deep tissue live imaging in post-implantation embryos and revealed an intriguing nuclear YAP pattern in migrating cells. The YAP fusion reporter mice and imaging methods will open new opportunities for understanding dynamic YAP signalling in vivo in many different situations.
Highlights
Yes-associated protein 1 (Yap1)–-commonly referred to as YAP—is a signalling protein that serves as a hub of biochemical and mechanical sensing [1,2,3]
YAP is a transcriptional co-activator for the TEA domain (TEAD) family transcription factors [6]
In response to numerous signals, YAP shifts its sub-cellular distribution to the nucleus where it associates with TEAD proteins to activate gene expression
Summary
Yes-associated protein 1 (Yap1)–-commonly referred to as YAP—is a signalling protein that serves as a hub of biochemical and mechanical sensing [1,2,3]. Real-time tracking of the dynamic YAP signalling status in vivo would offer crucial insights into the fundamental regulatory mechanisms of mammalian development and disease. In response to numerous signals, YAP shifts its sub-cellular distribution to the nucleus where it associates with TEAD proteins to activate gene expression. Monitoring the nuclear/cytoplasmic localization of YAP is the most broadly applied method to evaluate YAP activity [6,7]. This is primarily achieved by immunostaining of fixed embryos or tissue sections, precluding the acquisition of dynamic information. We report a YAP fusion reporter mouse line that allows in vivo live imaging of YAP behaviour
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