LIFE SCIENCE

This “gene scissors” alters cockroach genes


Researchers have developed a new method of CRISPR-Cas9 that makes cockroach gene editing possible. The results were recently published in the open science journal Cell Reporting Methods.

This simple and effective technique is named “direct parenting” CRISPR (DIPA-CRISPR). The technique injects the substance into the adult female whose egg is developing, rather than into the embryo itself.

“In a sense, insect researchers get rid of the hassle of injecting eggs.” Corresponding author Takaaki Daimon of Kyoto University in Japan said, “We can now edit insect genomes more freely and freely. In principle, this method should be suitable for more than 90% of insect species. ”

Currently, insect gene-editing methods often require the injection of material into early embryos, which severely limits its use in many species. For example, due to the unique reproductive system of cockroaches, past studies have failed to achieve genetic manipulation of them.

In addition, insect gene editing often requires expensive equipment, specific experimental devices for each species, and highly skilled laboratory personnel. “Problems with traditional methods plague researchers who want to edit the genomes of a wide variety of insects.” Daimon said.

To overcome these limitations, Daimon and collaborators injected Cas9 ribonucleoprotein (RNPs) into the main cavity of adult female cockroaches, causing genetic mutations in developing egg cells. The results show that the gene editing efficiency (the proportion of edited individuals to the total number of incubated individuals) can be as high as 22%. In the Chizhigu bandits, the efficiency of DIPA-CRISPR reaches more than 50%. In addition, the researchers produced beetle gene knock-ins by combining injections of single-stranded oligonucleotides and Cas9 RNPs, but the efficiency was low and needed to be further improved.

The researchers believe that the successful application of DIPA-CRISPR on two species with a longer evolutionary distance shows its potential for widespread application. But this method does not directly apply to all insects, including fruit flies.

In addition, experiments have shown that the most critical factor for success is the adult female who is injected. Therefore, DIPA-CRISPR requires a good understanding of insect ovarian development. Given the different life histories and reproductive strategies of insects, this can be a challenge for some species.

Despite these limitations, DIPA-CRISPR is accessible, highly practical, and can be easily implemented in the lab, extending gene editing applications to a wide range of model and non-model insect species. The technology requires only small devices for injection, and only two components, cas9 protein and monoconducting RNA, greatly simplifying the process of gene editing. In addition, commercialized standard Cas9 can be used for adult insect injections, eliminating the need for time-consuming protein customization engineering.

Courtesy of the author

“By improving the DIPA-CRISPR approach to make it more effective and versatile, we may be able to edit the genomes of almost all 1.5 million insect species, creating a future where we can take full advantage of the amazing biological functions of insects.” Daimon said, “In principle, it is also possible to edit the genomes of other arthropods using similar methods, including agricultural and medical pests such as mites and ticks, as well as important fishery resources such as shrimp and crabs.” (Source: China Science Daily Feng Weiwei)

Related paper information:https://doi.org/10.1016/j.crmeth.2022.100215



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