Spatio-temporal visualisation and manipulation of endogenous Yap1 in medaka
Seleit, A.; Asaoka, Y.; Knoblich, S.; Lederer, L.; Oozono, C.; Oga, A.; Oiso, A.; Furutani-Seiki, M.; Aulehla, A.
Show abstract
A key requirement to address fundamental questions in developmental biology is the ability to visualise and manipulate endogenous signalling proteins in vivo. Here we generate such an enabling experimental tool for the highly conserved Hippo pathway and its transcriptional effector Yap1, which converts mechanical and chemical cues into transcriptional output during embryogenesis and regeneration. We generate and characterise a Yap1- mGreenLantern (Yap1-mGL) endogenous CRISPR/Cas9-mediated knock-in line that enables 4D analysis of Yap1 dynamics in medaka. We resolve Yap1- mGL expression and nuclear/cytoplasmic localisation in real time, revealing the onset of reporter fluorescence before gastrulation, tissue and cell-type specific Yap1-mGL localisation during somitogenesis and injury-induced Yap1 upregulation during larval spinal cord regeneration. Using the nanobody-based degron system, we degrade the Yap1 protein in vivo, recapitulating the genetic loss-of-function mutant phenotype. Combining endogenous tags with nanobody-based degrons offers a powerful approach to visualise and acutely perturb endogenous protein function in vivo.
Matching journals
The top 3 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Degron tagging to label membrane-wrapped objects and probe membrane topology 95%
- A genetically encoded probe for imaging HA-tagged protein translation, localization, and dynamics in living cells and animals 94%
- Quantitative live imaging reveals PRICKLE1 controls junctional neural tube morphogenesis independent of Planar Cell Polarity 94%
Similar papers in this journal
Similar papers in this journal
Similar papers in this journal
- Self-reporting transposons enable simultaneous readout of gene expression and transcription factor binding in single cells 93%
- Latent developmental potential to form limb-like skeletal structures in zebrafish 93%
- NeuroPAL: A Neuronal Polychromatic Atlas of Landmarks for Whole-Brain Imaging in C. elegans 93%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.