Cellular functions depend on the functionality of proteins, and these functions are disturbed in diseases. A core aim of cell biological research is to determine the functions of individual proteins and how their disturbances result in disease. One way to study protein functions is to examine the effects of rapidly removing them from cells. During the past years, researchers have developed several techniques to achieve this. One of these techniques is knowing as AID, or auxin-inducible degron.
This method utilizes the signaling of a class of plant hormones known as auxins to rapidly deplete individual proteins from cells. The research group headed by Academy Professor Elina Ikonen at the University of Helsinki increased the speed and improved the hormone-dependency of the AID technique in human cells. The researchers were able to degrade the targeted cellular proteins within minutes. In addition, the researchers expanded the potential uses of the technique to encompass several types of proteins.
Functionality of proteins
The method can also be employing in the acute degradation of proteins whose long-term absence cannot be tolerating by cells. The technique we have developed is useful primarily in research, but thanks to advances in gene technology it also has potential for novel diagnostic and therapeutic methods. Auxin-inducible degron technology allows rapid and controlled protein depletion. However, basal degradation without auxin and inefficient auxin-inducible depletion have limited its utility.
We have identified a potent auxin-inducible degron system composed of auxin receptor F-box protein AtAFB2 and short degron miniIAA7. The system showed minimal basal degradation and enabled rapid auxin-inducible depletion of endogenous human transmembrane; cytoplasmic and nuclear proteins in 1 h with robust functional phenotypes. Perhaps the most important basic component of all cells are proteins that perform a wide variety of functions in cells and tissues.
In order to clarify the physiological roles of proteins, they are often linked to a green fluorescent protein (GFP) via targeted genetic manipulation, which makes them visible under the microscope. The observation of such GFP-linked proteins in living cells allows initial conclusions about the function of the protein. However, the exact function of a protein can often only be determining; when the protein is removing and the resulting consequences become visible in cells, tissues or model organisms.
But it was showing that the AID-nanobody can also be successfully using in the model organism zebrafish. Using the AID-nanobody in zebrafish demonstrating; for the first time that an auxin-mediated protein knockdown can also be implementing in a complex vertebrate model. In which different naturally occurring principles such as fluorescent GFP from algae; auxin-dependent protein degradation from plants; and the nanobody from camelids are combining to answer previously inaccessible research questions.