Optogenetic control of protein activity is a versatile technique to gain control over cellular processes, for example, for biomedical and biotechnological applications. Among other techniques, the regulation of protein abundance by controlling either transcription or protein stability found common use as this controls the activity of any type of target protein. Here, we report modules of an improved variant of the photosensitive degron module and a light-sensitive transcription factor, which we compared to doxycyclinedependent transcriptional control. Given their modularity the combined control of synthesis and stability of a given target protein resulted in the synergistic down regulation of its abundance by light. This combined module exhibits very high switching ratios, profound downregulation of protein abundance at low lightfluxes, and fast protein depletion kinetics. Overall, this synergistic optogenetic multistep control (SOMCo) module is easy to implement and results in a regulation of protein abundance superior to each individual component.
Synthetic
tools for the control of protein function are valuable
for biomedical research to characterize cellular functions of essential
proteins or if a rapid switch in protein activity is necessary. The
ability to tune protein activity precisely opens another level of
investigations that is not available with gene deletion mutants. Control
of protein stability is a versatile approach to influence the activity
of a target protein by its cellular abundance. Diverse strategies
have been developed to achieve efficient proteolysis using external
inducers or differentiation-coupled signals. The latter is especially
important for the inactivation of a protein during a developmental
process. Recently, several approaches to achieve this have been engineered.
In this article, we present current synthetic tools for regulation
of protein stability that allow fine-tuning of protein abundance,
their advantages and disadvantages with an emphasis on methods applicable
in the context of cell differentiation and development. We give an
outlook toward future developments and discuss main applications of
these tools.
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