CRISPR/Cas9 with egg electroporation
A quick way to create transgenic mice has been found
Geneticists from the University of California have improved the CRISPR/Cas9 genome editing technology. The new method is called CRISPR-EZ, and it allows you to insert new DNA molecules into the genomes of mouse embryos with almost one hundred percent success. The results of the study are published in the Journal of Biological Chemistry (Chen et al., Highly Efficient Mouse Genome Editing by CRISPR Ribonucleoprotein Electroporation of Zygotes).
The essence of the new approach is the use of electroporation. In order for the CRISPR/Cas9 system to be able to make changes to DNA, it is injected through a microscopic needle into fertilized eggs. A small electric discharge makes it possible to make a thin hole in the membrane through which the molecules of the system enter the cell.
In their experiment, the scientists managed to introduce a mutation into the gene in 88 percent of mice. This number exceeds the number of genetically modified mice obtained using the CRISPR-editing method, which uses conventional injections. According to scientists, their development can significantly improve the viability of embryos, which sometimes die due to genetic manipulation or are born with abnormalities.
CRISPR-EZ allows you to reduce the effort and time spent on obtaining mice with knocked out (defective) genes.
In vitro fertilization is used to create transgenic animals. First, the female is prepared for mating with the help of hormonal injections, after which scientists extract the fertilized eggs. The CRISPR/Cas9 system is injected into them through a thin needle. It consists of two types of RNA molecules, informational RNA, which encodes the cas9 protein, and guide RNA, which connects to a section of the mouse gene and indicates cas9 where to make a DNA cut.
Figure from the UC Berkeley Faster, more efficient CRISPR editing in mice press release – VM.
Portal "Eternal youth" http://vechnayamolodost.ru
30.05.2016