October 10, 2016

Editing the Epigenome in Mammals

What is the epigenome ?

The epigenome is a multitude of chemical compounds that can tell the genome what to do. The human genome is the complete assembly of DNA (deoxyribonucleic acid)-about 3 billion base pairs - that makes each individual unique. DNA holds the instructions for building the proteins that carry out a variety of functions in a cell. The epigenome is made up of chemical compounds and proteins that can attach to DNA and direct such actions as turning genes on or off, controlling the production of proteins in particular cells.

When epigenomic compounds attach to DNA and modify its function, they are said to have "marked" the genome. These marks do not change the sequence of the DNA. Rather, they change the way cells use the DNA's instructions. The marks are sometimes passed on from cell to cell as cells divide. They also can be passed down from one generation to the next.

What does the epigenome do?

A human being has trillions of cells, specialized for different functions in muscles, bones and the brain, and each of these cells carries essentially the same genome in its nucleus. The differences among cells are determined by how and when different sets of genes are turned on or off in various kinds of cells. Specialized cells in the eye turn on genes that make proteins that can detect light, while specialized cells in red blood cells make proteins that carry oxygen from the air to the rest of the body. The epigenome controls many of these changes to the genome.

What makes up the epigenome?

The epigenome is the set of chemical modifications to the DNA and DNA-associated proteins in the cell, which alter gene expression, and are heritable (via meiosis and mitosis). The modifications occur as a natural process of development and tissue differentiation, and can be altered in response to environmental exposures or disease.

The first type of mark, called DNA methylation, directly affects the DNA in a genome. In this process, proteins attach chemical tags called methyl groups to the bases of the DNA molecule in specific places. The methyl groups turn genes on or off by affecting interactions between the DNA and other proteins. In this way, cells can remember which genes are on or off.

The second kind of mark, called histone modification, affects DNA indirectly. DNA in cells is wrapped around histone proteins, which form spool-like structures that enable DNA's very long molecules to be wound up neatly into chromosomes inside the cell nucleus. Proteins can attach a variety of chemical tags to histones. Other proteins in cells can detect these tags and determine whether that region of DNA should be used or ignored in that cell.

Is the epigenome inherited?

The genome is passed from parents to their offspring and from cells, when they divide, to their next generation. Much of the epigenome is reset when parents pass their genomes to their offspring; however, under some circumstances, some of the chemical tags on the DNA and histones of eggs and sperm may be passed on to the next generation. When cells divide, often much of the epigenome is passed on to the next generation of cells, helping the cells remain specialized.

Highlights -Editing the Epigenome in Mammals

• dCas9-Tet1 and -Dnmt3a enable precise editing of CpG methylation in vitro and in vivo
• Targeted demethylation of BDNF promoter IV activates BDNF in neurons
• Targeted enhancer demethylation facilitates MyoD-induced muscle cell reprogramming
• Targeted de novo methylation of CTCF motifs alters CTCF-mediated gene loops

Summary -Editing the Epigenome in Mammals

Mammalian DNA methylation is a critical epigenetic mechanism orchestrating gene expression networks in many biological processes. However, investigation of the functions of specific methylation events remains challenging. Here, we demonstrate that fusion of Tet1 or Dnmt3a with a catalytically inactive Cas9 (dCas9) enables targeted DNA methylation editing. Targeting of the dCas9-Tet1 or -Dnmt3a fusion protein to methylated or unmethylated promoter sequences caused activation or silencing, respectively, of an endogenous reporter. Targeted demethylation of the BDNF promoter IV or the MyoD distal enhancer by dCas9-Tet1 induced BDNF expression in post-mitotic neurons or activated MyoD facilitating reprogramming of fibroblasts into myoblasts, respectively. Targeted de novo methylation of a CTCF loop anchor site by dCas9-Dnmt3a blocked CTCF binding and interfered with DNA looping, causing altered gene expression in the neighboring loop. Finally, we show that these tools can edit DNA methylation in mice, demonstrating their wide utility for functional studies of epigenetic regulation.

Journal Cell - Editing DNA Methylation in the Mammalian Genome

SOURCES- Journal Cell, Genome.gov

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