Back to Search Start Over

High‐throughput, microscopy‐based screening and quantification of genetic elements

Authors :
Rongrong Zhang
Yajia Huang
Mei Li
Lei Wang
Bing Li
Aiguo Xia
Ye Li
Shuai Yang
Fan Jin
Source :
mLife, Vol 2, Iss 4, Pp 450-461 (2023)
Publication Year :
2023
Publisher :
Wiley, 2023.

Abstract

Abstract Synthetic biology relies on the screening and quantification of genetic components to assemble sophisticated gene circuits with specific functions. Microscopy is a powerful tool for characterizing complex cellular phenotypes with increasing spatial and temporal resolution to library screening of genetic elements. Microscopy‐based assays are powerful tools for characterizing cellular phenotypes with spatial and temporal resolution and can be applied to large‐scale samples for library screening of genetic elements. However, strategies for high‐throughput microscopy experiments remain limited. Here, we present a high‐throughput, microscopy‐based platform that can simultaneously complete the preparation of an 8 × 12‐well agarose pad plate, allowing for the screening of 96 independent strains or experimental conditions in a single experiment. Using this platform, we screened a library of natural intrinsic promoters from Pseudomonas aeruginosa and identified a small subset of robust promoters that drives stable levels of gene expression under varying growth conditions. Additionally, the platform allowed for single‐cell measurement of genetic elements over time, enabling the identification of complex and dynamic phenotypes to map genotype in high throughput. We expected that the platform could be employed to accelerate the identification and characterization of genetic elements in various biological systems, as well as to understand the relationship between cellular phenotypes and internal states, including genotypes and gene expression programs.

Details

Language :
English
ISSN :
2770100X
Volume :
2
Issue :
4
Database :
Directory of Open Access Journals
Journal :
mLife
Publication Type :
Academic Journal
Accession number :
edsdoj.21f12515f24747119dd1548341552373
Document Type :
article
Full Text :
https://doi.org/10.1002/mlf2.12096