Fungal genome editing services, including knockout, knock-in, etc

Fungi, plural fungi, are any of the approximately 144,000 known organisms in the fungus kingdom, including yeast, rust, smut, mold, mold, and mushrooms. There are many types of fungi, including slime molds and oomycetes (water fungus), which do not belong to the fungus kingdom, but are usually called fungi. Many of these fungal-like organisms are included in the kingdom of Chromista. Fungi are one of the most widely distributed organisms on the earth, and are of great significance to the environment and medicine. Many fungi can live freely in the soil or water; other animals form a parasitic or symbiotic relationship with plants or animals, resulting in a decline in the somatic immunity of many animals and plants. It can be seen from science that gene knockout lines technology is very useful for suppressing knockout. Bacteria of various animals and plants.

Fungal pathogens are the main cause of the most serious diseases affecting plants, leading to a significant decline in yield and crop quality, and causing huge economic losses on a global scale. It is estimated that about 30% of emerging diseases are caused by fungi (Giraud et al., 2010), so new strategies are needed to improve their management. CRISPR-Cas9 genome editing technology (including gene knockout, knock-in, point mutation, etc.) (clustered regularly spaced short palindrome repeats-CRISPR-associated protein 9) allows researchers to modify the genome sequence more precisely. the way. The use of CRISPR-Cas not only provides a time-saving method for performing genome function analysis, but also provides new fungal genotypes that can be used as plant pathogens and/or potential competitors to trigger plant defense responses. By using CRISPR-Cas9, the activation of unknown clusters in beneficial fungi can be induced, thereby discovering new secondary metabolites that can interact with plants or plant pathogens. This can lead to the release of new and interesting biological control strains on site, avoid introducing transgenes into the environment, and gene knockout technology can well knock out Lentivirus Packaging caused by fungi.

Application of CRISPR/Cas9 in yeast: a useful genetic modification tool

The advancement that prompted many organisms to adopt CRISPR–Cas9 is its ability to target specific sites in the genome: generally, attempts to manipulate DNA fragments using homologous fragments of DNA are very inefficient. There is an exception in Saccharomyces cerevisiae, where gene editing can be achieved by converting sequences of less than 40 nucleotides (separated on both sides of the selectable marker) into cells. The highly homologous targeting of foreign DNA to the Saccharomyces cerevisiae genome is one of the reasons why yeast has become such a powerful experimental tool. Indeed, soon after its genome sequence was completed, every gene in the organism was deleted, and these mutants were available to researchers. Similarly, through homologous integration, all proteins are labeled with green fluorescent protein to understand the location of subcellular proteins, and epitopes are labeled to facilitate the overall analysis of protein complexes. Therefore, the gene editing system has far exceeded the tools used for genetic modification.

Controlling the occurrence of FHB by CRISPR/Cas9 silent mutationOne possible use of CRISPR-Cas9 silent mutants is Fusarium wilt (FHB), which is one of the most destructive diseases caused by different Fusarium species in global cereal crops, among which Fusarium graminearum and Fusarium bulbs. most. Ordinary and aggressive agents. In FHB, although the yield loss is due to the sterility of the infected florets, the decline in grain quality is mainly due to the accumulation of colistin (encoded by the fungal three-gene cluster) which is highly toxic to humans and animals. A previous study reported that the iRNA (interfering RNA) Δtri6 mutant of durum wheat showed that the disease index of durum wheat was reduced by 40% to 80% (Scherm et al., 2011). In addition, the classic knockout Δtri5 and Δtri6 mutants of Fusarium fusarium cannot spread the disease to adjacent spikelets and grains on wheat and corn, respectively, and cannot induce plant defense responses (Ravensdale et al., 2014). Similarly, the Δmap1 mutant of Fusarium fusarium showed a two-fold reduction in mycotoxin production, failed to produce membranes and could not penetrate wheat tissue, and did not affect the ability of straw colonization (Urban et al., 2003). . Considering that space and nutrient competition between virulent and non-virulent strains can reduce the disease, the on-site release of non-virulent CRISPR mutant strains of Fusarium graminearum and Fusarium subtilis may help control FHB. occur.

Gene knockout accelerates functional genomics research of Chlamydia Pseudomonas

Gene knockout technology is a useful molecular tool for studying gene function. However, chlamydia can resist a series of high levels.

Ubigene Biosciences is co-founded by biological academics and elites from China, the United States, and France. We are located in Guangzhou Science City, which serves as a global center for high technology and innovation. Ubigene Biosciences has 1000㎡ office areas and laboratories, involving genome editing, cell biology technology, and zebrafish research. We provide products and services for plasmids, viruses, cells, and zebrafish. We aim to provide customers with better gene-editing tools for cell or animal research.

We developed CRISPR-U™ and CRISPR-B™(based on CRISPR/Cas9 technology) which is more efficient than general CRISPR/Cas9 in double-strand breaking, CRISPR-U™ and CRISPR-B™ can greatly improve the efficiency of homologous recombination, easily achieve knockout (KO), point mutation (PM) and knockin (KI) in vitro and in vivo. 

References:

1. Nødvig CS, Hoof JB, Kogle ME, et al. Efficient oligonucleotide mediated CRISPR-Cas9 gene editing in Aspergilli. Fungal Genet Biol. 2018;115:78-89.

2.  Nødvig CS, Nielsen JB, Kogle ME, Mortensen, UH. A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015;10(7):e0133085. Published 2015 Jul 15.

3. Vanegas, K.G., Jarczynska, Z.D., Strucko, T. et al. Cpf1 enables fast and efficient genome editing in Aspergilli. Fungal Biol Biotechnol 6, 6 (2019).

4. Nayak T, Szewczyk E, Oakley CE, et al. A versatile and efficient gene-targeting system for Aspergillus nidulans. Genetics. 2006;172(3):1557-1566.