Synthetic biomolecular condensates to engineer eukaryotic cells

Area of Science:

• Cell Biology
• Synthetic Biology
• Biochemistry

Background:

• Eukaryotic cells compartmentalize functions using organelles.
• Non-membrane-bound biomolecular condensates offer synthetic biology potential.
• Controlling condensate formation is key for engineering cellular functions.

Purpose of the Study:

• To engineer mammalian cells with multiple, distinct genetic codes.
• To achieve precise protein engineering within living cells using orthogonal translation.
• To develop a strategy for spatially orthogonal enzyme engineering.

Main Methods:

• Designing multiple orthogonally translating organelles.
• Utilizing biomolecular condensates for spatial control.
• Engineering mammalian cells with expanded genetic codes.

Main Results:

• Achieved subresolution precision in placing engineered components within cells.
• Created mammalian cells with multiple, distinct genetic codes.
• Demonstrated the ability to engineer multiple proteins with distinct functionalities.

Conclusions:

• Orthogonal translation within biomolecular condensates enables precise spatial control of protein engineering.
• This approach provides a pathway for engineering complex functionalities in living eukaryotic cells.
• The strategy offers a general method for spatially orthogonal enzyme engineering.