Genomics Technologies for Cell Engineering

Abstract

The orchestrated action of genes controls cell type specifications, yet the systematic discovery of gene combinations for cell engineering 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. This technology platform is applicable to a broad range of biological settings, and will enable the systematic identification of genetic combinations that engineer specific cell types.