Fat – it’s vital for life, but too much can lead to a host of health problems. Studying how fatty or fatty tissue works in the body is essential for understanding obesity and other issues, but the structural differences in fat cells and their distribution throughout the body make this difficult.
“Fat cells are different from other cells in that they lack unique cell surface receptors and are only a minority of cells in fat tissue,” said Steven Romanelli, Ph.D., former lab member d’Ormand MacDougald, Ph. .D., Department of Molecular and Integrative Physiology.
In a new article published in the Journal of Biological Chemistry, Romanelli, MacDougald and their colleagues describe a breakthrough using CRISPR-Cas9, a tool that transformed research into molecular biology, but whose use in the study of adipose tissue was elusive.
“The biggest fat research challenge to date has been that if you want to study the function of a gene, you have to spend a considerable amount of time, resources and money developing a transgenic mouse.” , said Romanelli.
The traditional way to develop mouse models is to breed mice with a desired mutation to remove or introduce certain genes of interest, which Romanelli says can take over a year and tens of thousands of dollars.
CRISPR-Cas9 has revolutionized this process. This is a gene-editing technique made up of an enzyme called Cas9 that can break DNA strands and a piece of RNA that guides the Cas9 enzyme to a specific site in the genome for it. ‘editing. This tool is packaged in a harmless virus for delivery to the cells of interest. The tool has been used successfully to study the heart, liver, neurons, and skin cells to name a few, but never a certain type of fat cell known as brown fat.
Using this technique, the team successfully targeted brown fat, a specialized fatty tissue used to generate heat and protect core body temperature.
“What we were able to do was take this whole process and distill it in two weeks to a month to generate a transgenic mouse, reducing the cost to under $ 2,000. Not only does this reduce time and cost, but it democratizes research so that any laboratory familiar with molecular biology techniques can adopt this method and do it themselves, ”said Romanelli.
They were also able to use this method to suppress multiple genes simultaneously, a fact that could help researchers better understand important molecular pathways.
Using their CRISPR-Cas9 adeno-associated virus components, they successfully deactivated the UCP1 gene, which defines brown adipose tissue and allows it to generate heat, in adult mice. They observed that knockout mice were able to adapt to the loss of the gene and maintain body temperature in cold weather, hinting at other pathways involved in temperature homeostasis.
Romanelli says these early results are exploratory, but the technique represents an important step forward in the study of fat.
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Material provided by Michigan Medicine – University of Michigan. Original written by Kelly Malcom. Note: Content can be changed for style and length.