The mini-auxin inducible degree (mAID) system is a protein-targeted degradation technology based on the phytohormone growth hormone signaling pathway. By fusing a short degron tag to the target protein and co-expressing plant-derived F-box proteins in the cell, ubiquitination degradation of the target protein can be rapidly induced upon the addition of growth hormone. The system is characterized by a low basal degradation rate and high induction efficiency, which can achieve protein-specific depletion within a few hours. It provides a new tool for studying protein function with spatiotemporal dynamic control. In recent years, the mAID system has been widely used in eukaryotic organisms, especially in the study of gene function, construction of disease models, and validation of drug targets, showing significant advantages.

Lifeasible focuses on cutting-edge biotechnology applications and research. Based on mAID, we can help researchers perform precise and controllable protein modifications to rapidly knockdown the target proteins, etc., and have further research on the functions of target proteins in cell regulation and other aspects.

Features of mAID System?

• Time-sensitive. Protein modification can be completed within minutes, effectively capturing instantaneous changes in protein function.
• Highly controllable. By adjusting the concentration of growth hormone analogs, protein degradation can be "switched on and off."
• Reversible. After removing the inducer, intracellular protein expression can be gradually restored, which is helpful for dynamic functional studies.

Our Service for Knockdown Protein Using the mAlD System

mAID plasmid synthesis

The mAID system is derived from the mechanism of growth hormone-induced degradation in plants by fusing a miniature AID tag to the target protein, enabling rapid degradation of the target protein upon the addition of growth hormone analogs. We have extensive experience constructing a mAID plasmid that can efficiently express the fusion protein in cells, and we need to select the appropriate vector backbone, mAID tag sequence, and so on when constructing the mAID plasmid that can efficiently express the fusion protein. Below is our synthesis process and key steps.

• Sequence design and synthesis. Based on the gene sequence of the target protein and the expected fusion site, design the fusion gene containing the mAID tag. After the design is completed, the gene is synthesized or amplified by PCR for sequence validation.
• Plasmid backbone selection and preparation. Select a plasmid vector that is optimized and suitable for expression in mammalian cells. We perform restriction endonuclease digestion of the vector for subsequent insertion of the target fragment.
• Cloning of fusion fragments. The synthesized mAID fusion our target gene fragment is ligated to the digested vector.
• Bacterial transformation and screening. The ligation products are transformed into E. coli receptor cells and positive clones are obtained by resistance screening, followed by confirmation of sequence correctness.
• Plasmid extraction and quality control. In this step, we performed plasmid extraction and subsequent stringent quality control.

Plasmid synthesis

We use gene editing techniques to customize gene mAID tag insertion schemes to construct conditional protein degradation cell lines or nematode lines. Our gene editing plasmid construction mainly includes nuclease and sgRNA expression modules. After sgRNA design, nuclease expression vector construction, ligation reaction, bacteriophage transformation, positive screening, plasmid extraction, and validation, we complete the gene editing plasmid synthesis.

The combination of gene editing technology and the mAID system allows us to realize multi-level functional resolution from gene to protein. In addition, mAID-tagged cell lines constructed using gene editing technology can rapidly verify the gene function deletion phenotype, avoiding the irreversible limitations of traditional knockdown technology.

Our Service Workflow

• Project design and contracting. Provide customized technical solutions and sign service agreements.
• Experimental operation and model construction. We carry out gene editing, cell culture, and model construction according to the experimental program and provide real-time feedback on the experimental progress.
• Induction experiment and effect verification. Optimize the induction and degradation conditions to complete the rapid knockdown of the target protein and verify its effect from multiple angles.
• Organize data and technical reports. Organize experimental data, write detailed reports, and provide follow-up discussions and technical support on the results.

Fig.1 Our service using the mAlD system. (Lifeasible)

Why Choose Us?

• Rigorous quality control. We ensure that each plasmid synthesis and experimental step undergoes stringent quality assurance measures to ensure reliability and reproducibility.
• Customization and flexibility. Our team of experts works closely with customers to customize systems for specific research needs, providing tailored solutions that fit unique experimental parameters.
• Fast turnaround and expert support. From project conception to data analysis, our streamlined workflow and experienced staff provide timely and comprehensive support to help clients accelerate their studies.

Lifeasible's mAlD system provides advanced technology for protein knockdown and related research, as it enables efficient and reliable knockdown of the target proteins. If you are interested, please contact us.