Rapid and efficient degradation of endogenous proteins in vivo identifies stage-specific roles of RNA Pol II pausing in mammalian development.
Dev Cell
; 57(8): 1068-1080.e6, 2022 04 25.
Article
in En
| MEDLINE
| ID: mdl-35421370
ABSTRACT
Targeted protein degradation methods offer a unique avenue to assess a protein's function in a variety of model systems. Recently, these approaches have been applied to mammalian cell culture models, enabling unprecedented temporal control of protein function. However, the efficacy of these systems at the tissue and organismal levels in vivo is not well established. Here, we tested the functionality of the degradation tag (dTAG) degron system in mammalian development. We generated a homozygous knock-in mouse with a FKBP12F36V tag fused to negative elongation factor b (Nelfb) locus, a ubiquitously expressed regulator of transcription. In our validation of targeted endogenous protein degradation across mammalian development and adulthood, we demonstrate that irrespective of the route of administration the dTAG system is non-toxic, rapid, and efficient in embryos from the zygote-to-mid-gestation stages. Additionally, acute depletion of NELFB revealed a specific role in zygote-to-2-cell development and zygotic genome activation (ZGA).
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
RNA Polymerase II
/
Gene Expression Regulation, Developmental
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Dev Cell
Year:
2022
Document type:
Article