Disclosed herein include methods, compositions, and kits suitable for use in detecting the activation level of a signal transducer. In some embodiments, there are provided synthetic protein circuits wherein recruitment of synthetic protein circuit components to an association location upon activation of a signal transducer generates an active effector protein. The effector protein can be configured to carry out a variety of functions when in an active state, such as, for example, inducing cell death. Methods of treating a disease or disorder characterized by aberrant signaling are provided in some embodiments.

Disclosed herein include methods, compositions, and kits suitable for use in detecting the activation level of a signal transducer. In some embodiments, there are provided synthetic protein circuits wherein recruitment of synthetic protein circuit components to an association location upon activation of a signal transducer generates an active effector protein. The effector protein can be configured to carry out a variety of functions when in an active state, such as, for example, inducing cell death. Methods of treating a disease or disorder characterized by aberrant signaling are provided in some embodiments.

The present invention relates to synthetic α-secretase (SAS) and a use thereof. According to the present invention, a synthetic α-secretase, which is a fusion protein comprising, as an active ingredient, NIa protease, a fragment thereof or a variant thereof, can inhibit the formation of amyloid β, degrade extracellularly secreted amyloid β, and degrade amyloid β internalized in cells. Therefore, the present invention allows intracellular/extracellular degradation of amyloid β, which is the cause of various diseases including Alzheimer's disease, and thus can be usable in the prevention or treatment of such diseases.

The invention relates to a fusion protein containing a selective cell death-inducing enzyme system for use in the therapy and/or treatment of cancer and tumors in humans and animals, a process, and its use.

Provided are fusion proteins and related molecules useful for conducting base editing with reduced or no off-target mutations. The fusion protein may include a first fragment comprising a nucleobase deaminase or a catalytic domain thereof, a second fragment comprising a nucleobase deaminase inhibitor, and a protease cleavage site between the first fragment and the second fragment. Also provided are improved prime editing systems, including prime editing guide RNA with improved stability.

An insulin degludec derivative and a preparation method therefor are provided. Specifically, a fusion protein has a green fluorescent protein folding unit and an insulin degludec precursor or an active fragment thereof are provided. The fusion protein has a significantly increased expression level, and the insulin degludec precursor protein in the fusion protein is folded correctly, and has biological activity. Moreover, the green fluorescent protein folding unit in the fusion protein can be digested into small fragments by a protease, which have a great difference in molecular weight in comparison to a target protein, and are easy to separate. A method for using the fusion protein to prepare insulin degludec and prepare an intermediate thereof are also provided.

Tobacco etch virus protease (TEV) is one of the most widely used proteases in biotechnology because of its exquisite sequence-specificity. A limitation of TEV is its slow catalytic rate, which limits product generation and therefore signal output. Provided is a generalizable yeast-based platform for directed evolution of protease catalytic properties. Protease activity is determined via proteolytic release of a membrane-anchored transcription factor, and access to TEV's cleavage site is temporally regulated using a photosensory LOV domain. By gradually decreasing light exposure time, faster variants of TEV were selected over multiple rounds of selection. The mutant TEV proteases and the directed evolution platform are useful in a wide range of biotechnology applications, such as FLARE and SPARK tools.

To program intercellular communication for biomedicine, it is crucial to regulate the secretion and surface display of signaling proteins. If such regulations are at the protein level, there are additional advantages, including compact delivery and direct interactions with endogenous signalling pathways. A modular, generalizable design is provided called Retained Endoplasmic Cleavable Secretion (RELEASE), with engineered proteins retained in the endoplasmic reticulum and displayed/secreted in response to specific proteases. The design allows functional regulation of multiple synthetic and natural proteins by synthetic protease circuits to realize diverse signal processing capabilities, including logic operation and threshold tuning. By linking RELEASE to additional novel sensing and processing circuits, one would be able to achieve elevated protein secretion in response to “undruggable” oncogene KRAS mutants. RELEASE enables the local, programmable delivery of intercellular cues for a broad variety of fields such as neurobiology, cancer immunotherapy and cell transplantation.

Disclosed herein include methods and compositions for making proteins in peroxisomes as well as methods of making cells for producing proteins in peroxisomes. Also disclosed herein are cells for producing a protein in a peroxisome, and methods for producing a protein in a eukaryotic cell containing a peroxisome as described herein.

Aspects of the present disclosure are directed to biosensing circuits that correct crosstalk.