Posters
« Back
Potent transcriptional activation using CRISPRa with synthetic crRNA
EP26744
Poster Title: Potent transcriptional activation using CRISPRa with synthetic crRNA
Submitted on 07 Dec 2017
Author(s): Eldon T. Chou, Žaklina Strezoska, Elena Maksimova, Maren M. Gross, Melissa L. Kelley, and Anja van Brabant Smith
Affiliations: Dharmacon part of Horizon Discovery Group
This poster was presented at ASCB/EMBO Conference
Poster Views: 1,022
View poster »


Poster Information
Abstract: For CRISPR activation (CRISPRa), the guide RNA forms a complex with a nuclease-deactivated Cas9 (dCas9, D10A and H840A), which is in turn fused to transcriptional activators. The machinery then acts upstream of the transcription start site to up-regulate expression of a target gene. CRISPRa provides new tools to identify gene functions that might otherwise go undetected using loss-of-function studies through down-regulation of gene expression or gene knockout. Here we demonstrate a strategy to conduct CRISPRa experiments using chemically synthesized crRNA and tracrRNA molecules. We examine the functionality and advantages of using the synthetic approach for CRISPRa in multiple cell lines. Additionally, we show how the transcriptional activation leads to a significant increase in the target protein level, which in turn causes phenotypic effects by inhibiting or activating downstream genes. The methods presented are broadly applicable as a strategy to up-regulate any gene including systematic functional gene analysis in arrayed screening format.Summary: The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 (CRISPR-associated 9 proteins) system derived from Streptococcus pyogenes has been adapted to up-regulate any gene in its endogenous context, enabling gain-of-function or gene activation experiments while avoiding the use of exogenous over-expression plasmids. References: Report abuse »
Creative Commons

Related Posters


Cytotoxicity of Environmental Toxins PFOA and Gen X
Lauren Zane, Thomas Schultz

automated, low-cost, miniaturized RNA-Seqand DNAseqlibrary preps
SrimeenakshiSrinivasan2, PaymanehMalihi3, Peter De Hoff2, Tiffany Herrero2, To Cuong2, Joby Jenkins1, Louise Laurent2, James Hicks3, Peter Kuhn3

low-volume workflows improve the quality of drug discovery: two case studies
Margaret Kenney#, Archi Joardar#, Vasudeo Badarinarayana#, Joby Jenkins*

miniaturization and automation of CEL-Seq2 and SMART-Seq2 using the mosquito liquid handler
Josip Herman1, Jon Penterman2, Sagar1, Andreas Diefenbach3, Antigoni Triantafyllopoulou4, Anne F. Hammerstein5, Joby Jenkins5, Dominic Grün1, Stuart S. Levine2 and Klaus Hentrich5

ExpiSf™ Expression system: A Chemically Defined Baculovirus-Based System for Enhanced Protein and Virus Production in Sf9 Cells
Kenneth Thompson, Maya Yovcheva, Sara Barnes, Melissa Cross, Katy Irvin, Natasha Lucki, Henry Chiou, and Jonathan Zmuda