Saxena Pratik, Heng Boon Chin, Bai Peng, Folcher Marc, Zulewski Henryk, Fussenegger Martin
Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058 Basel, Switzerland.
Division of Endocrinology, Diabetes and Metabolism, University Hospital Basel, Petersgraben 4, CH-4031 Basel, Switzerland.
Nat Commun. 2016 Apr 11;7:11247. doi: 10.1038/ncomms11247.
Synthetic biology has advanced the design of standardized transcription control devices that programme cellular behaviour. By coupling synthetic signalling cascade- and transcription factor-based gene switches with reverse and differential sensitivity to the licensed food additive vanillic acid, we designed a synthetic lineage-control network combining vanillic acid-triggered mutually exclusive expression switches for the transcription factors Ngn3 (neurogenin 3; OFF-ON-OFF) and Pdx1 (pancreatic and duodenal homeobox 1; ON-OFF-ON) with the concomitant induction of MafA (V-maf musculoaponeurotic fibrosarcoma oncogene homologue A; OFF-ON). This designer network consisting of different network topologies orchestrating the timely control of transgenic and genomic Ngn3, Pdx1 and MafA variants is able to programme human induced pluripotent stem cells (hIPSCs)-derived pancreatic progenitor cells into glucose-sensitive insulin-secreting beta-like cells, whose glucose-stimulated insulin-release dynamics are comparable to human pancreatic islets. Synthetic lineage-control networks may provide the missing link to genetically programme somatic cells into autologous cell phenotypes for regenerative medicine.
合成生物学推动了标准化转录控制装置的设计，这些装置可对细胞行为进行编程。通过将基于合成信号级联和转录因子的基因开关与对许可食品添加剂香草酸具有反向和差异敏感性相结合，我们设计了一个合成谱系控制网络，该网络将香草酸触发的转录因子Ngn3（神经生成素3；关-开-关）和Pdx1（胰腺和十二指肠同源盒1；开-关-开）的互斥表达开关与MafA（V-maf肌腱膜纤维肉瘤癌基因同源物A；关-开）的伴随诱导相结合。这个由不同网络拓扑结构组成的设计网络能够协调对转基因和基因组Ngn3、Pdx1和MafA变体的适时控制，从而将人诱导多能干细胞（hIPSC）来源的胰腺祖细胞编程为对葡萄糖敏感的胰岛素分泌β样细胞，其葡萄糖刺激的胰岛素释放动力学与人类胰岛相当。合成谱系控制网络可能为将体细胞基因编程为用于再生医学的自体细胞表型提供缺失的环节。
