Dharmacon - RNAi, Gene Expression & Gene Editing

WEBINAR: CRISPRa tools for transcriptional activation studies

The CRISPR-Cas9 system has been adapted to upregulate any gene in its endogenous context, enabling overexpression experiments and avoiding the use of exogenous expression plasmids. The simplicity of programming this CRISPR activation (CRISPRa) system with small RNA guides is transformative for performing systematic gain-of-function studies. This exciting complement to loss-of-function studies (such as with RNAi and CRISPR-Cas9) also provides new tools to identify gene functions that might otherwise go undetected using a downregulation or knockout mechanism alone.

With CRISPRa, the guide RNAs form a complex with a nuclease-deactivated Cas9 (dCas9) which is in turn fused to transcriptional activators. This machinery then acts at the transcription start sites to upregulate expression of a target gene. In this webinar, we will describe different CRISPRa technologies and provide experimental considerations for using either synthetic guide RNA (crRNA:tracrRNA) or lentiviral sgRNA, in either arrayed or pooled screening formats. These platforms can be applied in tandem to enable both rapid follow-up of screen hits and development of complementary assays for functional gene analysis.

In this webinar you will learn:

  • The mechanisms of CRISPR activation
  • How to set up CRISPRa experiments using predesigned guide RNA, including delivery optimization and guide RNA format selection recommendations
  • The relationship between baseline gene expression and activation levels
  • The basics of using pooled lentiviral CRISPRa screening for target ID and validation

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