Institute of Zoology, CAS
研究员
中国科学院动物研究所
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, or the recently discovered RNA-targeting CRISPR systems.
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
New modules for DNA demethylation > Casilio-ME
Publications
Cheng, A.W.*#, Jillette, N.*, Lee, P., Plaskon, D., Fujiwara, Y., Wang, W., Taghbalout, A., Wang, H.# (2016) Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cell Research 26:254–257. doi: 10.1038/cr.2016.3 PMID:26768771
Taghbalout, A., Du, M., Jillette, N., Rosikiewicz, W. Rath, A., Heinen, C., Li, S., Cheng, A.W. (2019) Enhanced CRISPR-based DNA demethylation by Casilio-ME-mediated RNA-guided coupling of methylcytosine oxidation and DNA repair pathways. Nature Communications 10:4296. doi:10.1038/s41467-019-12339-7 PMID: 31541098 PMCID: PMC6754513
*Co-first; #Co-corresponding.
Selectable markers are limited in choices. We have created a collection of split selectable markers each allowing selection of multiple transgenes with one selection scheme, expanding the capacity of selectable markers. Read more
Publication:
Jillette, N., Du, M., Zhu, J.J., Cardoz, P., Cheng, A.W. (2019) Split Selectable Markers. Nature Communications 10:4968. doi: 10.1038/s41467-019-12891-2 PMID: 31672965 PMCID: PMC6823381
CRISPR Artificial Splicing Factors are RNA-guided splicing modulators based on dCasRx fusion with splicing factor proteins or domain thereof. See more
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.