西亚试剂:Genome-scale transcriptional activation by an engineered CR

发布时间：2025-12-03

Systematic interrogation of gene function requires the ability to perturb gene expression in a robust and generalizable manner. Here we describe structure-guided engineering of a CRISPR-Cas9 complex to mediate efficient transcriptional activation at endogenous genomic loci. We used these engineered Cas9 activation complexes to investigate single-guide RNA (sgRNA) targeting rules for effective transcriptional activation， to demonstrate multiplexed activation of ten genes simultaneously， and to upregulate long intergenic non-coding RNA (lincRNA) transcripts. We also synthesized a library consisting of 70，290 guides targeting all human RefSeq coding isoforms to screen for genes that， upon activation， confer resistance to a BRAF inhibitor. The top hits included genes previously shown to be able to confer resistance， and novel candidates were validated using individual sgRNA and complementary DNA overexpression. A gene signature based on the top screening hits correlated with a gene expression signature of BRAF inhibitor resistance in cell lines and patient-derived samples. These results collectively demonstrate the potential of Cas9-based activators as a powerful genetic perturbation technology.

 

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