Fats are a vital component of life but an excessive intake of them can lead to a host of health problems. Studying how adipose tissue or body fat functions is critical to understanding obesity and other issues.
But the structural differences in fat cells and their distribution throughout the body make this difficult to do.
“Fat cells differ from other cells in that they lack unique cell surface receptors and represent only a minority of cells within fat tissue,” said Stephen Romanelli, PhD, of the university’s Department of Molecular Physiology and Integration. Michigan.
In a new research paper published in the Journal of Biological Chemistry, Romanelli, Ormand MacDougald, Ph.D. Their colleagues described a breakthrough using CRISPR-Cas9, a tool that has transformed molecular biological research, but whose use in studying adipose tissue has been elusive.
It is a gene-editing technology that consists of an enzyme called Cas9, which can break strands of DNA, and a piece of RNA that directs the Cas9 enzyme to a specific site in the genome for editing. The tool has been used successfully to study heart cells, liver, nerve cells, and skin, to name a few, but a specific type of fat cell known as brown fat has not been used.
Using this technology, the team was able to successfully target brown fat, a specialized fatty tissue used to generate heat and protect core body temperature.
Using adenovirus-related CRISPR-Cas9 components, they knocked out the UCP1 gene that defines brown fat and enables thermogenesis in adult mice. They observed that the out-of-strength mice were able to adapt to the loss of the gene and maintain their body temperature in cold conditions, hinting at other pathways involved in temperature homeostasis.
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“The biggest challenge with lipid research so far is that if you want to study gene function, you have to devote a significant amount of time, resources and money to developing a transgenic mouse,” Romanelli said.
The traditional method for developing mouse models involves breeding mice with a desired mutation to delete or introduce specific genes of interest, which can take over a year and tens of thousands of dollars.
CRISPR-Cas9 technology has revolutionized this process.
“What we have been able to do is take this whole process and distill it anywhere from two weeks to a month to generate a genetically modified mouse, which brings the cost down to less than $2000. Not only does it reduce time and cost, it democratizes the research so that anyone can A laboratory familiar with molecular biology techniques could adopt this method and do it themselves.”
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They were also able to use this method to delete multiple genes simultaneously, a fact that could help researchers better understand important molecular pathways.
Although these results are exploratory, the breakthrough represents an important step forward in the study of lipids.
(Source: Michigan Medicine, University of Michigan – Posted by Kelly Malcolm)
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