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Researchers at Karolinska Institute and Stockholm University have developed a new technique that can simultaneously probe several different histone markers
in a single cell and thousands of cells.
This new method allows for a more detailed study of how cells in mouse brains acquire unique properties and specialization
.
The study was published in the journal Nature Biotechnology
.
Deciphering cellular diversity in the brain by simultaneously probing different epigenetic information in a single cell: Nano CUT & Tag uses modified nanobodies, antibody fragments that appear in camels such as alpacas, to map histone modifications
in thousands of individual cells in mouse brains.
"This technique is a stepping stone to understanding how cells transition between different states," says
Marek Bartosovic, a postdoctoral researcher at the Department of Medical Biochemistry and Biophysics at Karolinska Institute and now head of the research team at Stockholm University.
The cells in our bodies may be very different, but they inherit common genetic information
in the form of DNA in the genome.
To acquire different traits and specializations, cells read and interpret this common genome
in different ways.
One of the mechanisms for reading DNA involves a protein called histone, which binds
to the genome.
Histones exhibit variable surface markers and carry multiple layers of epigenetic information
.
These landmark histone modifications exist
in different patterns in the genomes of different cell types.
This allows each cell to interpret genomic information in different ways and then acquire different properties
.
Recently, several research teams, including the Karolinska Institutet team, have developed methods such as single-cell CUT & Tag, which allow the observation of individual histone modifications
at the single-cell level and over a large scale.
Now, the Karolinska Institutet research team has taken the leap further by developing a new technology, Nano CUT & Tag
.
The technology is based on a new type of molecule
called nanobodies.
Nanobodies are small proteins that recognize other proteins like antibodies, are highly specific, but smaller and can easily fuse
with other proteins.
Using different nanobodies fused with an enzyme called Tn5 transposase allows different histone modifications
to be probed simultaneously in the same cell.
Nano CUT & Tag provides unique insights into how cells interpret genomes to specialize and gain a unique identity
by simultaneously tweaking multiple layers of epigenetic information.
"Nano CUT & Tag, or Nano-CT, allowed us to dissect in great detail how progenitor cells in the brain specialize into oligodendrocytes, a cell type that is the target of an autoimmune attack in multiple sclerosis," said
Goncalo Castelo-Branco, a professor of glial cell biology at Karolinska Institutet.
"These new mechanistic insights could provide clues
as to how we stimulate the recovery of oligodendrocytes in the context of disease.
"
Marek Bartosovic, who recently set up his research group at Stockholm University and will focus on the development of single-cell epigenomic technologies in the context of early human brain development, said: "We now want to further develop Nano-CT by increasing the range
of epigenetic markers it can detect.
"
