Researchers have succeeded in editing the genome of black-footed mites using CRISPR-Cas9. To achieve this feat, they have developed an embryonic injection protocol that overcomes the major barriers in the field.The work will be published in the journal on February 15th. iScience..
“Despite the ability to acquire and inherit a range of debilitating pathogens, mite studies are primarily available genetic and Molecular tool“Mr. Monika Gulia-Nuss, co-chief author of the study and molecular biologist at the University of Nevada, Reno.”
“By making genome editing tools available, Tick We will create a genome that will enable us to determine how these unique animals survive in the environment, interact with pathogens, and prevent mites from spreading the disease to humans and livestock. ” She says.
Despite the public health implications of mites, which are carriers of a wide variety of pathogens for humans, wildlife and livestock, knowledge of mite biology at the molecular level is still limited. This is in contrast to insects such as mosquitoes, for which numerous tools for transgenic development and genome editing are currently available. “Advances in this area are important for advances in research to solve the growing problem of tick-borne diseases,” said another co-author of the study and an entomologist at the University of Nevada, Reno. One Andrew Nus says.
CRISPR-Cas9 has revolutionized functional genetics research in many organisms. This gene editing technique has been applied to mediators of mosquitoes and other arthropod diseases, but successful gene editing in mites has not yet been reported.Technical challenges for injecting mites Embryo Attempts to edit genes have further slowed the progress of research. Tick embryos are very difficult to inject for the following reasons: High pressure A wax layer on the inside of the egg, the hard chorion (the outer shell of the egg), and the outside of the embryo. These should be removed before injection. Female mites use a special organ called the Gene’s organ to cover their eggs with a tough wax layer.
In iScience In the study, researchers developed a successful tick embryo injection protocol and target gene disruption in CRISPR-Cas9 using two methods: embryo injection and receptor-mediated cargo ovarian transduction (ReMOT control). Did. Arthropod. Researchers removed Gene’s organs to prevent wax deposition, and then treated the eggs with chemicals called benzalkonium chloride and sodium chloride to remove the chorion and reduce the pressure inside the eggs. ..
“We were able to carefully dissect pregnant female mites and surgically remove the organs responsible for waxing the eggs, but the females are still able to lay viable eggs. These are wax-free. egg “We allowed the materials needed for genomic modification to be injected into mite embryos,” said Gulia-Nuss. “Another major challenge was understanding the timing of mite embryogenesis. Little is known, so it was necessary to determine the exact time of introduction of CRISPR-Cas9 to maximize the potential for induction. Genetic changes.. “
The viability of the injected embryos is about 10%, comparable to the well-established insect model. In the case of ReMOT control, all injected mites survived.Data show the feasibility of mite embryo injection Genetic engineering Proboscipedia (with ticks by both methods that had the same editing efficiency of ProboscipediaProbP) Genes expressed in the appendages.
“Previously, no laboratory has demonstrated that mites can modify the genome. Some thought it would be too technically difficult to achieve this,” says Nuss. “This is the first study to demonstrate that genetic transformation of mites is possible in two different ways, not just one.”
Further research is needed to fully understand the molecular mechanisms underlying the efficient gene editing of mites. While these tools accelerate tick genetic research, improved embryo injection protocols are needed to increase viability and efficiency of larval hatching and gene editing.
“We hope that the tools we develop here will pave the way for new research that will dramatically accelerate our understanding of the molecular biology of this mite species and related mite species,” says Gulia-Nuss. increase. “Target disruption of genes in tick vectors of human pathogens reveals the underlying biology of tick-pathogen-host interactions that can inform the development and application of new approaches to tick-borne disease control. It’s a powerful method. ”
Monika Gulia-Nuss, Cas9-mediated gene editing, embryo injection and ReMOT control of the black-footed tick Ixodes scapularis iScience (2022). DOI: 10.1016 / j.isci.2022.103781.. www.cell.com/iscience/fulltext… 2589-0042 (22) 00051-7
Quote: Gene editing is now possible with Tick (February 15, 2022). Obtained from https: //phys.org/news/2022-02-gene.html on February 15, 2022
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Gene editing is now possible with ticks
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