Printing and Erasing of DNA‐Based Photoresists Inside Synthetic Cells

Abstract

In the pursuit of producing functioning synthetic cells from the bottom‐up, DNA nanotechnology has proven to be a powerful tool. However, the crowded yet highly organized arrangement in living cells, bridging from the nano‐ to the micron‐scale, remains challenging to recreate with DNA‐based architectures. Here, laser microprinting is established to print and erase shape‐controlled DNA hydrogels inside the confinement of water‐in‐oil droplets and giant unilamellar lipid vesicles (GUVs). The DNA‐based photoresist consists of a photocleavable inactive DNA linker, which interconnects Y‐shaped DNA motifs when activated by local irradiation with a 405 nm laser. An alternative linker design allows to erase custom features from a preformed DNA hydrogel with feature sizes down to 1.38 μm. The present work demonstrates that the DNA hydrogels can serve as an internal support to stabilize non‐spherical GUV shapes. Overall, DNA‐based photoresists for laser printing in confinement allow to build up architectures on the interior of synthetic cells with light, which diversifies the toolbox of bottom‐up synthetic biology.

Identifying and Manipulating Giant Vesicles: Review of Recent Approaches

“…Thus, through computational design, the construction of molecular aggregate in a 3D manner is practically and reproducibly controlled. Göpfrich’s group developed a two-photon laser-assisted 3D printing technology inside GUVs [ 115 , 116 ]. They constructed tubular 3D molds not only inside but also in the intramembrane of GUV and the membrane permeability of the GUV was changed by the mold.…”

DNA Droplets: Intelligent, Dynamic Fluid

“…DNA hydrogels, with sizes ranging from nanometer to centimeter scales, have a mesh-like 3D network structure with properties such as biodegradability, high permeability, biocompatibility, low elasticity, and stimuli-responsiveness. [30,49,[81][82][83][84][85] DNA hydrogels have witnessed various applications, such as cell-free protein production (Figure 2B), [86][87][88] pH sensing (Figure 2C), [89] DNA-based laser printing (Figure 2D), [90] photolithographic gel shape control, [91] multi-florescence emission (Figure 3B), [67] and dynamic soft material with flow-regulatable locomotion (Figure 3E). For a detailed description, refer to more comprehensive reviews of DNA hydrogels in the literature, [30,49,[81][82][83][84]92,93] together with some reviews for hydrogel in general.…”

“…Reproduced under the terms of the Creative Commons CC-BY license. [90] Copyright 2022, The Authors, published by John Wiley and Sons. (ii) Hydrogel property changes upon exposure to air and water.…”

“…Reproduced under the terms of the Creative Commons CC‐BY license. [ 90 ] Copyright 2022, The Authors, published by John Wiley and Sons. In “DNA nanostructures,” (DNA origami) Scale bar, 100 nm.…”

“…(Laser printing/erasing) Scale bar, 100 µm. Reproduced under the terms of the Creative Commons CC-BY license [90]. Copyright 2022, The Authors, published by John Wiley and Sons.…”