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Using Models to (Re-)Design Synthetic Circuits.

Author(s): McCallum G, Potvin-Trottier L

Mathematical models play an important role in the design of synthetic gene circuits, by guiding the choice of biological components and their assembly into novel gene networks. Here, we present a guide for biologists to build and utilize models of gene netw...

Article GUID: 33405217
Isolating live cells after high-throughput, long-term, time-lapse microscopy.

Author(s): Luro S, Potvin-Trottier L, Okumus B, Paulsson J

Nat Methods. 2019 Nov 25;: Authors: Luro S, Potvin-Trottier L, Okumus B, Paulsson J

Article GUID: 31768062
Bacterial variability in the mammalian gut captured by a single-cell synthetic oscillator.

Author(s): Riglar DT, Richmond DL, Potvin-Trottier L, Verdegaal AA, Naydich AD, Bakshi S, Leoncini E, Lyon LG, Paulsson J, Silver PA

Nat Commun. 2019 Oct 11;10(1):4665 Authors: Riglar DT, Richmond DL, Potvin-Trottier L, Verdegaal AA, Naydich AD, Bakshi S, Leoncini E, Lyon LG, Paulsson J, Silver PA

Article GUID: 31604953
Title:Isolating live cells after high-throughput, long-term, time-lapse microscopy.
Authors:Luro S,  Potvin-Trottier L,  Okumus B,  Paulsson J
Link:https://www.ncbi.nlm.nih.gov/pubmed/31768062?dopt=Abstract
DOI:10.1038/s41592-019-0620-7
Category:Nat Methods
PMID:31768062
Dept Affiliation: BIOLOGY
1 Department of Systems Biology, Harvard Medical School, Boston, MA, USA. spl53@cornell.edu.
2 Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
3 Department of Biology, Concordia University, Montreal, Québec, Canada.
4 Illumina, Foster City, CA, USA.
5 Department of Systems Biology, Harvard Medical School, Boston, MA, USA. johan_paulsson@hms.harvard.edu.

Description:

Isolating live cells after high-throughput, long-term, time-lapse microscopy.

Nat Methods. 2019 Nov 25;:

Authors: Luro S, Potvin-Trottier L, Okumus B, Paulsson J

Abstract

Single-cell genetic screens can be incredibly powerful, but current high-throughput platforms do not track dynamic processes, and even for non-dynamic properties they struggle to separate mutants of interest from phenotypic outliers of the wild-type population. Here we introduce SIFT, single-cell isolation following time-lapse imaging, to address these limitations. After imaging and tracking individual bacteria for tens of consecutive generations under tightly controlled growth conditions, cells of interest are isolated and propagated for downstream analysis, free of contamination and without genetic or physiological perturbations. This platform can characterize tens of thousands of cell lineages per day, making it possible to accurately screen complex phenotypes without the need for barcoding or genetic modifications. We applied SIFT to identify a set of ultraprecise synthetic gene oscillators, with circuit variants spanning a 30-fold range of average periods. This revealed novel design principles in synthetic biology and demonstrated the power of SIFT to reliably screen diverse dynamic phenotypes.

PMID: 31768062 [PubMed - as supplied by publisher]