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Engineering 'cell robots' for parallel and highly sensitive screening of biomolecules under in vivo conditions
Citation Link: https://doi.org/10.15480/882.1495
Publikationstyp
Journal Article
Publikationsdatum
2017-11-09
Sprache
English
Author
Enthalten in
Volume
7
Issue
1
Start Page
15145
Citation
Scientific reports 1 (7): 15145- (2017-11-09)
Publisher DOI
Scopus ID
Publisher
Nature Publishing Group
Cells are capable of rapid replication and performing tasks adaptively and ultra-sensitively and can be considered as cheap "biological-robots". Here we propose to engineer cells for screening biomolecules in parallel and with high sensitivity. Specifically, we place the biomolecule variants (library) on the bacterial phage M13. We then design cells to screen the library based on cell-phage interactions mediated by a specific intracellular signal change caused by the biomolecule of interest. For proof of concept, we used intracellular lysine concentration in E. coli as a signal to successfully screen variants of functional aspartate kinase III (AK-III) under in vivo conditions, a key enzyme in L-lysine biosynthesis which is strictly inhibited by L-lysine. Comparative studies with flow cytometry method failed to distinguish the wild-type from lysine resistance variants of AK-III, confirming a higher sensitivity of the method. It opens up a new and effective way of in vivo high-throughput screening for functional molecules and can be easily implemented at low costs.
Schlagworte
High-throughput screening
Metabolic engineering
DDC Class
570: Biowissenschaften, Biologie
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