(R)evolution-on-a-chip

Actinosomes: Condensate-Templated Containers for Engineering Synthetic Cells

“… 4 This has led to the design of a wide variety of confinements such as surfactant-stabilized water-in-oil droplets, liposomes with a lipid bilayer as the boundary, and even completely synthetic containers such as polymersomes and dendrimersomes. 5 , 6 These compartments are capable of reconstituting various biochemical processes within them and have been exploited to engineer a wide variety of cellular modules and to advance various applications like cell-free gene expression, 7 , 8 evolving proteins by directed evolution, 9 cytoskeleton assembly, 10 , 11 growth and division, 12 − 14 cargos for drug delivery, 15 and printing artificial tissues. 16 , 17 In these confinements formed via the hydrophobic effect, 18 the membrane usually acts as a physical barrier and restricts passive transport of molecules across them.…”

Actinosomes: condensate-templated proteinaceous containers for engineering synthetic cells

“…This has led to the design of a wide variety of confinements such as surfactant-stabilized water-in-oil droplets, liposomes with a lipid bilayer as the boundary, and even completely synthetic containers such as polymersomes and dendrimersomes [5,[7][8][9] . These compartments are capable of reconstituting various biochemical processes within them and have been exploited to engineer a wide variety of cellular modules and to advance various applications like cell-free gene expression [10,11] , evolving proteins by directed evolution [12] , cytoskeleton assembly [13,14] , growth and division [15][16][17][18][19] , cargos for drug delivery [20] , and printing artificial tissues [21,22] . In these confinements formed via the hydrophobic effect [23] , membrane usually acts as a physical barrier and restricts passive transport of molecules across them.…”

Domestication of proteins – from evolution to revolution

“…Hence, efficiencies should be improved at all levels: generation of genetic libraries, compartmentalisation, and last but not least smart screening or selection approaches. Especially major recent developments of both microfluidics technology and of high‐throughput automated sorting methods of cells or droplets (FACS/FADS) hold promise for unprecedented possibilities in the (near) future to obtain proteins with desired optimal features (reviewed by Bouzetos et al, 2021). As Charles Darwin stated: ‘There is grandeur in this view of life, (…) from so simple a beginning endless forms most beautiful and wonderful have been and are being evolved’.…”

“…To allow tracing a protein variant with a desired functionality back to its gene, a genotype‐to‐phenotype linkage is a key requirement. This can be achieved either by physically linking the gene and gene‐encoded product (DNA display, mRNA display, ribosome display), or by compartmentalising the gene and the corresponding protein within the same physical space (reviewed by Bouzetos et al, 2021). Unicellular microorganisms (e.g.…”

“…In IVC, single gene variants of a library are engulfed in artificial compartments such as water‐in‐oil droplets, or water‐in‐oil‐in‐water droplets. Recent progress in microfluidic technology has allowed for the production of highly monodisperse droplets (reviewed by Bouzetos et al, 2021). Gene expression inside these artificial compartment is catalysed by in vitro transcription and translation systems.…”