Tran Mai P, Chatterjee Rakesh, Dreher Yannik, Fichtler Julius, Jahnke Kevin, Hilbert Lennart, Zaburdaev Vasily, Göpfrich Kerstin
Biophysical Engineering Group, Max Planck Institute for Medical Research, Jahnstr. 29, 69120, Heidelberg, Germany.
Department of Biosciences, Heidelberg University, 69120, Heidelberg, Germany.
Small. 2023 Mar;19(13):e2202711. doi: 10.1002/smll.202202711. Epub 2022 Aug 15.
The bottom-up construction of an artificial cell requires the realization of synthetic cell division. Significant progress has been made toward reliable compartment division, yet mechanisms to segregate the DNA-encoded informational content are still in their infancy. Herein, droplets of DNA Y-motifs are formed by liquid-liquid phase separation. DNA droplet segregation is obtained by cleaving the linking component between two populations of DNA Y-motifs. In addition to enzymatic cleavage, photolabile sites are introduced for spatio-temporally controlled DNA segregation in bulk as well as in cell-sized water-in-oil droplets and giant unilamellar lipid vesicles (GUVs). Notably, the segregation process is slower in confinement than in bulk. The ionic strength of the solution and the nucleobase sequences are employed to regulate the segregation dynamics. The experimental results are corroborated in a lattice-based theoretical model which mimics the interactions between the DNA Y-motif populations. Altogether, engineered DNA droplets, reconstituted in GUVs, can represent a strategy toward a DNA segregation module within bottom-up assembled synthetic cells.
人工细胞的自下而上构建需要实现合成细胞分裂。在可靠的区室分裂方面已经取得了重大进展，但分离DNA编码信息内容的机制仍处于起步阶段。在此，DNA Y基序液滴通过液-液相分离形成。通过切割两个DNA Y基序群体之间的连接成分来实现DNA液滴分离。除了酶切之外，还引入了光不稳定位点，用于在本体以及细胞大小的油包水乳滴和巨型单层脂质囊泡（GUV）中进行时空控制的DNA分离。值得注意的是，受限环境中的分离过程比本体中的要慢。利用溶液的离子强度和核碱基序列来调节分离动力学。在基于晶格的理论模型中证实了实验结果，该模型模拟了DNA Y基序群体之间的相互作用。总之，在GUV中重构的工程化DNA液滴可以代表一种在自下而上组装的合成细胞中实现DNA分离模块的策略。
