High-Multiplexed Genome Engineering Using Synthetic CRISPR Arrays

Genetic engineering of biological systems is a fundamental tool for both basic and translation research, where up- and down-regulation of gene expression is necessary to drive cellular phenotypes and evaluate gene function. However, modulating the expression of multiple genes is difficult and labor-intensive.

Researchers in Stanley Qi's laboratory at Stanford have developed a CRISPR-based tool for multiplexed genome engineering. The inventors have combined multiple Cas enzymes and developed guide sequences to drive multi-gene silencing and activation in mammalian cells. While existing CRISPR arrays are limited by interference between adjacent RNAs, the inventors have optimized linker sequences to minimize interference between guide RNA sequences.

Among its many applications, the technology can be used to control developmental transcriptional programs, and differentially modulating gene signatures related to immune response. It can be used with different delivery methods including transfection or transduction to aid cell engineering and manufacturing.