Adaptation of the AID system for stem cell and transgenic mouse research.

Stem Cell Research
Marina V PryzhkovaPhilip W Jordan

Abstract

The auxin-inducible degron (AID) system is becoming a widely used method for rapid and reversible degradation of target proteins. This system has been successfully used to study gene and protein functions in eukaryotic cells and common model organisms, such as nematode and fruit fly. To date, applications of the AID system in mammalian stem cell research are limited. Furthermore, standard mouse models harboring the AID system have not been established. Here we have explored the utility of the H11 safe-harbor locus for integration of the TIR1 transgene, an essential component of auxin-based protein degradation system. We have shown that the H11 locus can support constitutive and conditional TIR1 expression in mouse and human embryonic stem cells, as well as in mice. We demonstrate that the AID system can be successfully employed for rapid degradation of stable proteins in embryonic stem cells, which is crucial for investigation of protein functions in quickly changing environments, such as stem cell proliferation and differentiation. As embryonic stem cells possess unlimited proliferative capacity, differentiation potential, and can mimic organ development, we believe that these research tools will be an applicable resource to a...Continue Reading

Citations

Nov 23, 2021·Frontiers in Molecular Biosciences·Anastasia YunusovaNariman Battulin

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Methods Mentioned

BETA
transgenic
gene knockout
mechanical dissociation
transfection
PCR
genotyping
fluorescence microscopy

Software Mentioned

GraphPad Prism
GeneTools
BZ
Snapgene
Envigo
GenBank
ZEN
Photoshop
Zhang Lab
X800 Viewer and Analyzer

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