PASIV - A pooled approach-based workflow to overcome toxicity-induced Design of Experiments failures and inefficiencies
File(s)
Published version
Author(s)
Casas, Alexis
Bultelle, Matthieu
Motraghi, Charles
Kitney, Richard
Type
Journal Article
Abstract
We present here a newly developed workflow - which we have called PASIV - designed to provide a solution to a practical problem with Design of Experiments methodology (DoE): i.e. what can be done if the scoping phase of the DoE cycle is severely hampered by burden and toxicity issues (either caused by the metabolite or an intermediary) making it unreliable or impossible to proceed to the screening phase? PASIV - standing for Pooled Approach, Screening, Identification and Visualization - was designed so the (viable) region of interest can be made to appear through an interplay between biology and software. This was achieved by
combining multiplex construction in a pooled approach (one pot reaction) with a viability assay, and with a range of bioinformatic tools (including a novel construct matching tool). PASIV was tested on the exemplar of the lycopene pathway - under stressful constitutive expression yielding a region of interest with comparatively stronger producers.
combining multiplex construction in a pooled approach (one pot reaction) with a viability assay, and with a range of bioinformatic tools (including a novel construct matching tool). PASIV was tested on the exemplar of the lycopene pathway - under stressful constitutive expression yielding a region of interest with comparatively stronger producers.
Date Issued
2022-03-09
Date Acceptance
2022-02-07
Citation
ACS Synthetic Biology, 2022, 11 (3), pp.1272-1291
ISSN
2161-5063
Publisher
American Chemical Society
Start Page
1272
End Page
1291
Journal / Book Title
ACS Synthetic Biology
Volume
11
Issue
3
Sponsor
Engineering & Physical Science Research Council (EPSRC)
Identifier
https://pubs.acs.org/doi/10.1021/acssynbio.1c00562
Grant Number
EP/S001859/1
Subjects
construct matching
design of experiments
pooled approach
scoping
0304 Medicinal and Biomolecular Chemistry
0601 Biochemistry and Cell Biology
0903 Biomedical Engineering
Publication Status
Published
Date Publish Online
2022-03-09