Systematic Evaluation of CRISPRa and CRISPRi Modalities Enables Development of a Multiplexed, Orthogonal Gene Activation and Repression System.
Andrea MartellaMike FirthBenjamin J M TaylorAnne GöppertEmanuela M CuomoRobert G RothAlan J DicksonDavid I FisherPublished in: ACS synthetic biology (2019)
The ability to manipulate the expression of mammalian genes using synthetic transcription factors is highly desirable in both fields of basic research and industry for diverse applications, including stem cell reprogramming and differentiation, tissue engineering, and drug discovery. Orthogonal CRISPR systems can be used for simultaneous transcriptional upregulation of a subset of target genes while downregulating another subset, thus gaining control of gene regulatory networks, signaling pathways, and cellular processes whose activity depends on the expression of multiple genes. We have used a rapid and efficient modular cloning system to build and test in parallel diverse CRISPRa and CRISPRi systems and develop an efficient orthogonal gene regulation system for multiplexed and simultaneous up- and downregulation of endogenous target genes.
Keyphrases
- genome wide
- genome wide identification
- poor prognosis
- transcription factor
- stem cells
- drug discovery
- signaling pathway
- tissue engineering
- bioinformatics analysis
- dna methylation
- genome wide analysis
- cell proliferation
- single cell
- gene expression
- long non coding rna
- oxidative stress
- dna binding
- quantum dots
- endoplasmic reticulum stress
- heat shock protein
- loop mediated isothermal amplification
- cell therapy