Application of high-throughput techniques in the discovery of microbial resources and functional genes

Abstract:

In light of the continuous discovery of microbial resources and the accumulating microbial genome data, it is an urgent task to screen and identify functional genes associated with important microbial phenotypes from the enormous data, for which high-throughput techniques are indispensible. The techniques are mainly involved in library construction and screening. Library construction refers to the establishment of mutant or interference libraries covering the whole genome of target microorganisms with techniques such as metagenomics, transposon insertion, RNA interference (RNAi), retron library recombineering (RLR), CRISPR interference (CRISPRi), and CRISPR activation (CRISPRa). As for the screening, a certain stress is often used to allow the differential growth of microbial individuals of a mutant library and then the causal relationships between specific genes and phenotypic outcomes at genomic level are identified by high-throughput sequencing. In this review, we briefly summarize the current high-throughput techniques used in functional genomics research, hoping to provide a reference to the development and optimization of these techniques in the future.