Bacterial Microcolonies in Gel Beads for High-Throughput Screening of Libraries in Synthetic Biology.

Détails

Ressource 1Télécharger: Duarte_Barbier_Schaerli.pdf (903.91 [Ko])
Etat: Public
Version: de l'auteur⸱e
Document(s) secondaire(s)
Télécharger: Supporting_Information.pdf (469.66 [Ko])
Etat: Public
Version: de l'auteur⸱e
ID Serval
serval:BIB_6AE8F65F20EA
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Bacterial Microcolonies in Gel Beads for High-Throughput Screening of Libraries in Synthetic Biology.
Périodique
ACS synthetic biology
Auteur⸱e⸱s
Duarte J.M., Barbier I., Schaerli Y.
ISSN
2161-5063 (Electronic)
ISSN-L
2161-5063
Statut éditorial
Publié
Date de publication
17/11/2017
Peer-reviewed
Oui
Volume
6
Numéro
11
Pages
1988-1995
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
Synthetic biologists increasingly rely on directed evolution to optimize engineered biological systems. Applying an appropriate screening or selection method for identifying the potentially rare library members with the desired properties is a crucial step for success in these experiments. Special challenges include substantial cell-to-cell variability and the requirement to check multiple states (e.g., being ON or OFF depending on the input). Here, we present a high-throughput screening method that addresses these challenges. First, we encapsulate single bacteria into microfluidic agarose gel beads. After incubation, they harbor monoclonal bacterial microcolonies (e.g., expressing a synthetic construct) and can be sorted according their fluorescence by fluorescence activated cell sorting (FACS). We determine enrichment rates and demonstrate that we can measure the average fluorescent signals of microcolonies containing phenotypically heterogeneous cells, obviating the problem of cell-to-cell variability. Finally, we apply this method to sort a pBAD promoter library at ON and OFF states.

Mots-clé
cell-to-cell variability, combinatorial libraries, directed evolution, hydrogel beads, screening, synthetic biology
Pubmed
Web of science
Open Access
Oui
Création de la notice
09/11/2017 18:18
Dernière modification de la notice
20/08/2019 15:25
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