What are the advantages of DNA technology
News portal - Ruhr University Bochum
The Crispr technology has made changing the genetic material much easier. The advantages and disadvantages of the method are discussed by Privatdozent Dr. Thorsten Müller from the Ruhr University Bochum and Dr. Hassan Bukhari from Harvard Medical School in an overview article in the journal “Trends in Cell Biology” on September 12, 2019. They see potential for future research primarily in making the Crispr technique applicable to stem cells.
In order to investigate the effects of genes or to make gene products visible, they used to be artificially over-activated. "They were then up to a thousand times more common than naturally," says Thorsten Müller. "The cell was flooded with gene products, the proteins, which can falsify the analysis of the function." This disadvantage does not apply with the Crispr method. It can be used to smuggle blueprints for fluorescent proteins into cells and place them behind a specific gene. "This makes it possible for the first time to follow the function of a protein live under natural conditions - and not after a 1,000-fold overproduction," explains the biochemist.
The Crispr method itself has also evolved. For the first method, researchers had to create so-called vectors at great expense in order to mark genes in the genome. The vectors are DNA segments, the sequence of which must be partially identical to the DNA of the target cell so that the gene to be introduced finds the right place. Today the natural DNA repair function of the cells is used, which also significantly simplifies the creation of the vectors, and thus fluorescent proteins can be introduced quickly and easily.
How drugs work
The fluorescent labeling also makes it possible to observe live under the microscope where in the cell the labeled gene products are located. “That could be interesting, for example, to test the effects of drugs on certain gene products,” explains Müller. To do this, the researchers would have to stimulate the cell with the active ingredient and monitor whether or how the location of the gene product changes.
Different genes can be marked with different colored fluorescences and analyzed in parallel. The more a gene marked in this way is read, the more the cell fluoresces in the corresponding color.
Organoids could replace animal research
The authors see particular potential for the method in combination with so-called organoids. These are mini-organs made from induced pluripotent stem cells that can be obtained from an adult organism. This can be used, for example, to build mini-brains that functionally correspond to the human brain.
If the Crispr technology is increasingly applied to stem cells in the future, researchers could investigate the effects of gene modifications not only in isolated cells, but also in complex tissue structures. "We could examine human genes live in human-like tissues and would be much less dependent on animal models," concludes Müller.
Based on these considerations, Müller and Bukhari work out a number of central research questions in the article that would have to be answered in order to bring the Crispr technique and the Organoid technique together.
Hassan Bukhari, Thorsten Müller: Endogenous fluorescence tagging by CRISPR, in: Trends in Cell Biology, 2019, DOI: 10.1016 / j.tcb.2019.08.004
Privatdozent Dr. Thorsten Müller
Chair of Biochemistry II
Faculty of Chemistry and Biochemistry
Tel .: 0234 32 24404
Email: [email protected]
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