ArticleFrontiers in molecular biosciences2021
Evaluation of the OsTIR1 and AtAFB2 AID Systems for Genome Architectural Protein Degradation in Mammalian Cells.
Article in Frontiers in molecular biosciences, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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7 citing papers in PubMed, 19 citations in OpenAlex.
- Uniform dynamics of cohesin-mediated loop extrusion in living human cells.Nature genetics · 2025Article
- The Influence of Cohesin on the Short-Scale Dynamics of Intact and Damaged Chromatin in Different Phases of the Cell Cycle.International journal of molecular sciences · 2025Article
- Coregulation of NDC80 Complex Subunits Determines the Fidelity of the Spindle-Assembly Checkpoint and Mitosis.Molecular cancer research : MCR · 2024Article
- A robust dual gene ON-OFF toggle directed by two independent promoter-degron pairs.Journal of cell science · 2023Article
- Function and Evolution of the Loop Extrusion Machinery in Animals.International journal of molecular sciences · 2023Review
- The human EF1a promoter does not provide expression of the transgene in mice.Transgenic research · 2022Article
- A One-step strategy to target essential factors with auxin-inducible degron system in mouse embryonic stem cells.Frontiers in cell and developmental biology · 2022Article
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6 authors at 3 institutions in 1 country.
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Abstract
The auxin-inducible degron (AID) system is a promising tool for dynamic protein degradation. In mammalian cells, this approach has become indispensable to study fundamental molecular functions, such as replication, chromatin dynamics, or transcription, which are otherwise difficult to dissect. We present evaluation of the two prominent AID systems based on OsTIR1 and AtAFB2 auxin receptor F-box proteins (AFBs). We analyzed degradation dynamics of cohesin/condensin complex subunits in mouse embryonic stem cells (Rad21, Smc2, Ncaph, and Ncaph2) and human haploid HAP1 line (RAD21, SMC2). Double antibiotic selection helped achieve high homozygous AID tagging of an endogenous gene for all genes using CRISPR/Cas9. We found that the main challenge for successful protein degradation is obtaining cell clones with high and stable AFB expression levels due to the mosaic expression of AFBs. AFB expression from a transgene tends to decline with passages in the absence of constant antibiotic selection, preventing epigenetic silencing of a transgene, even at the
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