Advanced combinatorial genetics technology for decoding biological complexity

Abstract

The orchestrated action of genes controls complex biological traits, yet the systematic discovery of gene combinations that modulate these phenotypes in human cells is labor-intensive and challenging to scale. Harnessing the power of synthetic biology and next-generation sequencing technologies, we developed CombiGEM (Combinatorial Genetics En Masse) as a powerful platform for high-throughput functional characterization of combinatorial genetic perturbations in human cells. CombiGEM enables rapid, scalable assembly of high-order barcoded combinatorial genetic libraries, and multiplexed quantification of all library members by using next-generation sequencing technologies. The genetic elements included in CombiGEM libraries can be arbitrary, including microRNAs, gene expression/knockdown constructs, and programmable CRISPR-Cas genome editing tools. We envision that this technology platform will be applicable to a broad range of biological settings, and will enable the systematic identification of genetic combinations that regulate complex biological processes and diseases.