Quantitative Tracking of Combinatorially Engineered Populations with Multiplexed Binary Assemblies.
ACS Synth Biol
; 6(4): 619-627, 2017 04 21.
Article
em En
| MEDLINE
| ID: mdl-28103008
ABSTRACT
Advances in synthetic biology and genomics have enabled full-scale genome engineering efforts on laboratory time scales. However, the absence of sufficient approaches for mapping engineered genomes at system-wide scales onto performance has limited the adoption of more sophisticated algorithms for engineering complex biological systems. Here we report on the development and application of a robust approach to quantitatively map combinatorially engineered populations at scales up to several dozen target sites. This approach works by assembling genome engineered sites with cell-specific barcodes into a format compatible with high-throughput sequencing technologies. This approach, called barcoded-TRACE (bTRACE) was applied to assess E. coli populations engineered by recursive multiplex recombineering across both 6-target sites and 31-target sites. The 31-target library was then tracked throughout growth selections in the presence and absence of isopentenol (a potential next-generation biofuel). We also use the resolution of bTRACE to compare the influence of technical and biological noise on genome engineering efforts.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Algoritmos
/
Engenharia Genética
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article