High-Throughput Functional Genomics for Energy Production.

[Display omitted] • High-throughput (HTP) functional genomics enables complex production phenotypes. • CRISPR-Cas-mediated genome editing expands HTP functional genomic experiments. • Pathway and tolerance engineering require multiplex mutagenesis approaches. Functional genomics remains a foundational field for establishing genotype-phenotype relationships that enable strain engineering. High-throughput (HTP) methods accelerate the Design-Build-Test-Learn cycle that currently drives synthetic biology towards a forward engineering future. Trackable mutagenesis techniques including transposon insertion sequencing and CRISPR-Cas-mediated genome editing allow for rapid fitness profiling of a collection, or library, of mutants to discover beneficial mutations. Due to the relative speed of these experiments compared to adaptive evolution experiments, iterative rounds of mutagenesis can be implemented for next-generation metabolic engineering efforts to design complex production and tolerance phenotypes. Additionally, the expansion of these mutagenesis techniques to novel bacteria are opening up industrial microbes that show promise for establishing a bio-based economy. [ABSTRACT FROM AUTHOR]