FROM GENE EDITING TO EPIGENETIC EDITING

Removing the CAS "scissors"

Again, any homing enzyme can work, but Cas9 is the best. The 2 nuclease domains on Cas9, which normally cut DNA, can mutated to generate a "DEAD" CAS. It may be ”dead” for cutting, but GENOME HOMING is all good. 

Deliverying Epi-Modifying Cargo

gRNAs give Cas HOMING COORDINATES. Headed to an promoter? enhancer? Find out about choosing good targets among EPIGENOMIC DATA and designing gRNAs. Then just choose what's in the "cargo box." 

Choosing Epi-MOD Cargo

Targeting histones? Targeting DNA for methylation? There's an enzyme for that.

Connecting Epi-MOD Cargo to DNA homing "dead" Cas

Most connections are done through GENE FUSION. That is, the gene sequence for "dead" Cas9 is literally connected to a particular cargo of EPIGENETIC MODIFYING ENZYME through RECOMBINANT DNA TECHNOLOGY. Once a cell has this new "fusion" sequence, it will read and produce custom EPIGENETIC ENZYME. 

Testing for the Epi-Change

DNA methylation changes. Cardiomyocyte Differentiation. Tompkins et al, 2016

On the basics of detecting EPIGENETIC and EPIGENOMIC changes. How do you test for changes in HISTONE ACETYLATION, DNA METHYLATION, and other EPI-MODS? 

SLOW DOWN. TAKE ME BACK TO THE BASICS

READY FOR MORE. Check out these recent papers on advances in EPIGENOME ENGINEERING

Links to papers with more on epigenetic engineering

ON EPIGENETIC MECHANISMS AND EPIGENETIC ENGINEERING CONCEPTS


Epigenome Editing: State of the Art, Concepts, and Perspectives. Kungulovski and Jeltsch, 2016. Trends in Genetics. 

https://www.cell.com/trends/genetics/fulltext/S0168-9525(15)00212-7


Next stop for the CRISPR revolution: RNA‐guided epigenetic regulators. Vora et al, 2016. The FEBS journal.

https://febs.onlinelibrary.wiley.com/doi/full/10.1111/febs.13768


An epigenetic perspective on the failing heart and pluripotent-derived-cardiomyocytes for cell replacement therapy. Tompkins and Riggs, 2015. Frontiers in Biology. 

https://link.springer.com/article/10.1007%2Fs11515-014-1340-0


The epigenome: the next substrate for engineering. Park et al, 2016. Genome Biology. 

https://genomebiology.biomedcentral.com/articles/10.1186/s13059-016-1046-5


SPECIFIC EXAMPLES OF EPIGENETIC MANIPULATION PAPERS.


Editing DNA Methylation in the Mammalian Genome. Lui et al, 2016. Cell.

https://www.cell.com/cell/fulltext/S0092-8674(16)31153-9


Epigenome editing by a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Hilton et al, 2015. Nature Biotech.

https://www.nature.com/articles/nbt.3199


Functional annotation of native enhancers with a Cas9-histone demethylase fusion. Kearns et al, 2015. Nature Methods. 

https://www.nature.com/articles/nmeth.3325