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INTRODUCTION

The Cheng Lab develops technologies based on artificial DNA and RNA binding proteins for genome editing, epigenome editing and transcriptome editing. For DNA binding proteins, the main one used in the lab is CRISPR/Cas9. For RNA binding proteins, the main one is Pumilio.

CRISPR/Cas9

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and CRISPR-associated (Cas) …

Pumilio

Pumilio proteins are ..

Casilio = CRISPR/Cas + Pumilio

Casilio is a modular platform of DNA binding enzymes based on a hybrid of CRISPR/dCas9 and Pumilio-tetherd effectors.  It allows multiplexed and combinatorial operations at different targets in the same cell. Modularity of the platform allows new effector modules to be added seamlessly, expanding the functionality of CRISPR/Cas beyond genome editing. See more

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RESEARCH PROJECTS

  1. Improving precise genome editing
  2. Developing novel approaches for epigenetic and transcript editing
  3. Developing sensors for DNA and RNA
  4. Developing imaging techniques for 3D structure of the genome.

Dr. CHENG’s PAST RESEARCH

  • CRISPR-on

    An RNA-guided transcriptional activator

    Technologies allowing for specific regulation of endogenous genes are valuable for the study of gene functions and have great potential in therapeutics. We created the CRISPR-on system, a two-component transcriptional activator consisting of a nuclease-dead Cas9 (dCas9) protein fused with a transcriptional activation domain and single guide RNAs (sgRNAs) with complementary sequence to gene promoters. We demonstrate that CRISPR-on can efficiently activate exogenous reporter genes in both human and mouse cells in a tunable manner. In addition, we show that robust reporter gene activation in vivo can be achieved by injecting the system components into mouse zygotes. Furthermore, we show that CRISPR-on can activate the endogenous IL1RN, SOX2, and OCT4genes. The most efficient gene activation was achieved by clusters of 3-4 sgRNAs binding to the proximal promoters, suggesting their synergistic action in gene induction. Significantly, when sgRNAs targeting multiple genes were simultaneously introduced into cells, robust multiplexed endogenous gene activation was achieved. Genome-wide expression profiling demonstrated high specificity of the system.

    Cell Research (2013) 23:1163-1171

    http://crispr-on.org