TY - JOUR
T1 - In Vitro Transcription-Translation in an Artificial Biomolecular Condensate
AU - Schoenmakers, Ludo L J
AU - Yewdall, N Amy
AU - Lu, Tiemei
AU - André, Alain A M
AU - Nelissen, Frank H T
AU - Spruijt, Evan
AU - Huck, Wilhelm T S
PY - 2023/7/21
Y1 - 2023/7/21
N2 - Biomolecular condensates are a promising platform for synthetic cell formation and constitute a potential missing link between the chemical and cellular stage of the origins of life. However, it has proven challenging to integrate complex reaction networks into biomolecular condensates, such as a cell-free in vitro transcription-translation (IVTT) system. Integrating IVTT into biomolecular condensates successfully is one precondition for condensation-based synthetic cell formation. Moreover, it would provide a proof of concept that biomolecular condensates are in principle compatible with the central dogma, one of the hallmarks of cellular life. Here, we have systemically investigated the compatibility of eight different (bio)molecular condensates with IVTT incorporation. Of these eight candidates, we have found that a green fluorescent protein-labeled, intrinsically disordered cationic protein (GFP-K72) and single-stranded DNA (ssDNA) can form biomolecular condensates that are compatible with up to μM fluorescent protein expression. This shows that biomolecular condensates can indeed integrate complex reaction networks, confirming their use as synthetic cell platforms and hinting at a possible role in the origin of life.
AB - Biomolecular condensates are a promising platform for synthetic cell formation and constitute a potential missing link between the chemical and cellular stage of the origins of life. However, it has proven challenging to integrate complex reaction networks into biomolecular condensates, such as a cell-free in vitro transcription-translation (IVTT) system. Integrating IVTT into biomolecular condensates successfully is one precondition for condensation-based synthetic cell formation. Moreover, it would provide a proof of concept that biomolecular condensates are in principle compatible with the central dogma, one of the hallmarks of cellular life. Here, we have systemically investigated the compatibility of eight different (bio)molecular condensates with IVTT incorporation. Of these eight candidates, we have found that a green fluorescent protein-labeled, intrinsically disordered cationic protein (GFP-K72) and single-stranded DNA (ssDNA) can form biomolecular condensates that are compatible with up to μM fluorescent protein expression. This shows that biomolecular condensates can indeed integrate complex reaction networks, confirming their use as synthetic cell platforms and hinting at a possible role in the origin of life.
KW - Artificial Cells
KW - Biomolecular Condensates
KW - Coloring Agents
KW - DNA, Single-Stranded
KW - Green Fluorescent Proteins/genetics
KW - biomolecular condensation
KW - coacervation
KW - liquid−liquid phase separation (LLPS)
KW - in vitro transcription translation (IVTT)
KW - synthetic cell
UR - http://www.scopus.com/inward/record.url?scp=85164402654&partnerID=8YFLogxK
U2 - 10.1021/acssynbio.3c00069
DO - 10.1021/acssynbio.3c00069
M3 - Journal article
C2 - 37343188
SN - 2161-5063
VL - 12
SP - 2004
EP - 2014
JO - ACS Synthetic Biology
JF - ACS Synthetic Biology
IS - 7
ER -