Natural evolution programs drastic changes in cellular function through genetic changes to a cell’s genome. Both germline and somatic evolution simultaneously and combinatorially explore complex genetic changes across perturbation classes, including gene knockouts, knockdowns, overexpression, and the creation of new genes from existing domains. Separate bespoke technologies have been developed to perform each of these types of genetic perturbations at scale in human cells, but these methods are largely mutually incompatible. 

We have developed CRISPR-All, a unified genetic perturbation language for programming of any major type of genetic perturbation simultaneously, in any combination, at genome scale, in primary human cells. Analogous to computational programming languages, CRISPR-All converts high level descriptions of desired complex genetic changes into a single DNA sequence that can rewire genomic programs within a cell. 

CRISPR-All allows for comprehensive analysis of genetic enhancements, such as direct comparisons of all proposed human CAR-T cell functional enhancements from over 30 years of study simultaneously head to head. Arbitrarily multiplexed genome scale screens of combinatorial genetic perturbations can easily be designed, constructed, and run. And CRISPR-All is natively compatible with single cell sequencing, allowing high throughput linkage of complex, multiplexed genetic perturbation programs with high dimensional phenotypes in primary human cells. Across disease settings and human cell types, CRISPR-All enables exploration of a combinatorial genetic perturbation space similar to the genetic diversity accessed by natural evolution, allowing us to more fully explore basic evolutionary mechanisms and clinical applications of genetic perturbations. 

Genetically encoded therapies offer the promise for programmable, rapidly adaptable, and ultimately more powerful therapeutic cellular manipulations. We have applied successive generations of genetic discovery and gene design technologies to develop improved engineered human TCR and CAR T cell therapies for cancer immunotherapy. Yet the challenges posed by complex disease such as cancer, autoimmunity, neurodegeneration, and aging continue to resist current generations of genetic, as well as small molecule and biologic therapies. 

We continue to develop and apply ever more powerful genetic discovery tools to identify complex combinations of genetic manipulations, as well as new gene designs, to drive more powerful cellular therapies. We pair these input sequences with ever improving ex vivo and in vivo delivery modes across human cell types, with the long term aim to understand and therapeutically translate dynamic genetic therapies, ensuring the right sequence is expressed by the right cell at the right time to change the course of a patient’s complex disease.