44 results about "Gene drive" patented technology

The invention belongs to the field of bioengineering and particularly relates to a Gene drive carrier and a construction method thereof. The construction method includes the steps of 1), artificiallysynthesizing a cap5A promoter and an sgRNA fragment; 2), amplifying a cas9 gene; 3), amplifying a plasmid skeleton; 4), amplifying an rpsL promoter; 5), assembling an empty carrier; 6), inserting a spacer targeted at the formula described; 7), constructing the carrier. In the method, a novel chromosome box recombinase CcrC2 and CRISPR-Cas9 technology are combined in an attempting manner, an SCCmeckiller carrier system, namely the Gene driver carrier is constructed, the Gene drive acts on methicillin-resistant staphylococcus aureus MRSA, staphylococcal chromosome box SCCmec is targeted to remove from the methicillin-resistant staphylococcus aureus MRSA, and methicillin-resistant genes are thereby eliminated.

The present invention is related to the field of CRISPR-Cas9 gene editing platforms. In particular, the present invention has identified Type II-C Cas9 anti-CRISPR (Acr) inhibitors that control Cas9 gene editing activity. Co-administration of such Acr inhibitors may provide an advantageous adjunct in permitting safe and practical biological therapeutics through spatial or temporal control of Cas9activity; controlling Cas9-based gene drives in wild populations to reduce the ecological consequences of such forced inheritance schemes; and contributing to general research into various biotechnological, agricultural, and medical applications of gene editing technologies.

The invention discloses a plant inducible promoter and an application thereof. A section of HbCIPK2 promoter sequence of 1,750bp is obtained from a halophyte wild barley genome through a chromosome primer walking method, an arabidopsis proHbCIPK2:GUS strain obtained through the promoter sequence can drive expression of GUS genes under stress of different concentrations and times, and the expression activity of GUS is enhanced along with increase of the stress times and concentrations. The inducible expression activity of the GUS genes driven by the HbCIPK2 promoter under PEG and ABA stress is relatively low. The identical transgenic lines are free of GUS gene expressions under stress-free treatment and low-temperature and high-temperature stress. The obtained DNA sequence of 1,750 bp is the novel stress inducible promoter, and has high promoter activity under high-salt stress, and proper expressions of transcription factors and other controlling genes can be induced.

The invention relates to a root-specific promoter and application thereof, belonging to the field of a biological technology. A root-specific expression promoter has a nucleotide sequence shown by SEQ ID NO 1. The root-specific expression promoter is obtained by cloning at the root of peanut; and experiments prove that a gene driven by the promoter is specifically expressed at the roots of arabidopsis thaliana, tobacco and peanut. The root-specific expression promoter has important significance to prevention and control of diseases and pests of roots of plants.

The invention discloses a construction method for a transgene mouse model of overexpressing an Rps23r1 gene, belonging to transgene models, in particular relating to the construction method for the transgene mouse model of specifically over-expressing the Rps23r1 gene in a brain. A transgene mouse of over-expressing the Rps23r1 gene is obtained through cloning the Rps23r1 gene, constructing a recombinant plasmid and injecting into a germ cell of the mouse. A genome of the transgene mouse of over-expressing the Rps23r1 gene stably integrates the Rps23r1 gene driven by a mouse Prion gene promoter and can be stably inherited to offspring and express an exogenous RPS23R1 protein with an Myc mark in the brain. The Rps23r1 transgene mouse is found not to have abnormal changes in the long-term observation and provides effective means for researching and estimating Rps23r1 physiological and pathologic functions in vivo.

Described herein are embodiments relating to manipulation of populations and sex ratio in populations through DNA sequence modifications.

The invention belongs to the technical field of plant molecular biology, and in particular discloses a shepherd's purse peroxidase gene promoter and an application thereof in improving plant cold tolerance. The nucleotide sequence of the shepherd's purse peroxidase gene promoter is that of the 1st-1021st bases in SEQ ID No.1. A gene driven by the promoter can code a peroxidase gene which is inducible by low temperature and is capable of regulating reactive oxygen in plant cells. Under an induction condition of 4 DEG C, the expression amount of the promoter is up-regulated, and meanwhile, the expression amount of the promoter is also up-regulated in leaves and stems. An inducible plant expression vector, which is constructed by virtue of promoter fragments, can start the expression of a target gene in the case that plants are under low-temperature stress. The invention also provides an application of the promoter in breeding cold-tolerance plants, so that the cold tolerance of crop varieties can be improved.

The invention relates to a method for improving features of nano-crystalline cellulose by using a glycosyl transferase 8D1 promoter to drive a GA20ox. The method comprises the following steps: using the glycosyl transferase 8D1 promoter of the xylem specific expression to drive the gibberellin oxidase GA20ox gene to excessively express and change the features of the aspen wood nano-crystalline cellulose; through acquiring a recombinant vector excessively expressed by the GA20ox gene driven by the promoter 8D1P specifically expressed in the aspen stalk xylem, enabling the recombinant vector tobe used for genetic transformation of an aspen, to obtain the 8D1P:GA20ox transgene 84K aspen, and achieving a purpose of improving the features of the wood nano-crystalline cellulose. The improvementof the features of the wood nano-crystalline cellulose of the transgene aspen is that a diameter of the nano-crystalline cellulose is increased, and a draw ratio of the nano-crystalline cellulose isincreased. A fiber diameter of the nano-crystalline cellulose of a transgene material is larger, the mechanical strength of fibers can be remarkably improved, and the above feature change is more beneficial to applications of pulping and papermaking.

Described herein are embodiments relating to manipulation of populations and sex ratio in populations through DNA sequence modifications.

The present invention relates to a vaccine comprising the insertion of three genes, the TAA / ecdCD40L EA1 and CDA, driven by promoters L-plastin / cytosinedeaminase and CMV as a three gene, three transcription unit oncolytic virus as a conditionally replication competent adenoviral vector which replicates only in tumor cells. In these transcription units, the E1A gene of the adenoviral vector as well as the cytosine deaminase gene are under the control of the L-plastin promoter, while the TAA / ecdCD40L transcription unit is under control of a the CMV promoter.

The present invention is related to the field of CRISPR-Cas9 gene editing platforms. In particular, the present invention has identified Type II-C Cas9 anti-CRISPR (Acr) inhibitors that control Cas9 gene editing activity. Co-administration of such Acr inhibitors may provide an advantageous adjunct in permitting safe and practical biological therapeutics through spatial or temporal control of Cas9 activity; controlling Cas9-based gene drives in wild populations to reduce the ecological consequences of such forced inheritance schemes; and contributing to general research into various biotechnological, agricultural, and medical applications of gene editing technologies.

The invention belongs to the technical field of transgenosis and in particular discloses a preparation method of transgenic tilapia with a lysozyme gene. A new tilapia strain which can achieve induced expression of tilapia lysozyme C3 is obtained by introducing a Tgf2 transposon carrying a lysozyme C3 gene driven by an Hsp70 promoter into tilapia zygotes and integrating the Hsp70 promoter and the lysozyme C3 gene onto a tilapia genome by utilizing the transposition characteristic of the transposon. By adopting the preparation method, not only can the immunity of the tilapia be improved and the new tilapia variety with stronger disease resistance be bred but also the Tgf2 transposon is introduced into construction of the transgenic tilapia for the first time and a new method is provided for importing exogenous genes into cultured fishes. The preparation method has great theoretical significance and application value.

The invention provides a recombinant adenovirus features that it can simultaneously inactivate the virus proteins able to combine with cell internal funcational tumor inhibition gene p53 and inhibit cell decline, respectively, and carries with a group of exogenous genes driven by exogenous stator. It can prolifically breed in P53 function defect-type and / or P53 function normality-type tumor cells, generating a cytopathic effect (CPE), and can make high-performance expression exogenous antitumor proteins in the tumor cells, and finally kills the tumor cells, but has no killing effect on P53 function normality-type non-tumor cells.

Methods and compositions for improving the specificity of genome-editing nucleases (e.g., RNA-guided CRISPR-Cas nucleases or engineered zinc finger nucleases) and customizable DNA-binding domain fusion proteins (e.g., RNA-guided dead-Cas9, RNA-guided dead-Cpf1, or engineered zinc finger arrays fused to transcriptional regulatory domains) for use as research reagents, in gene drives, or as therapeutic agents.

The invention discloses a floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof, belonging to separation and application of a promoter. The nucleotide sequence of the promoter is shown as SEQ ID NO.1. The invention also discloses a recombinant plant expression vector and recombinant host cell containing the floral organ specific expression promoter. Functional verification tests discover that a GUS gene driven by the promoter only performs specific expression in sepals and petals of tobacco, and no GUS expression activity exists in roots, stems, leaves, stamens, carpels and other tissues of the tobacco, thus indicating that the promoter separated from Populus trichocarpa Torr.&Gray is the floral organ specific expression promoter. The invention further discloses application of the floral organ specific expression promoter in the aspects of improving plant traits, cultivating new plant species, studying floral organ development and regulation mechanisms of plants and the like.

The invention discloses application of a CRISPR / Cas9 genome editing system in alfalfa gene editing, the CRISPR / Cas9 genome editing system comprises an expression vector, the expression vector can comprise a Cas9 expression cassette, an sgRNA1 expression cassette and an sgRNA2 expression cassette, and the Cas9 expression cassette is used for expressing Cas9. The sgRNA1 expression cassette is used for expressing sgRNA with the name of sgRNA1, and the sgRNA2 expression cassette is used for expressing sgRNA with the name of sgRNA2; the sgRNA1 expression cassette contains a promoter with the name of MtU6-6promoter and a sgRNA1 gene driven by the MtU6-6promoter, and the sgRNA2 expression cassette contains a promoter with the name of MtU6-5promoter and a sgRNA2 gene driven by the MtU6-5promoter; the sgRNA1 and the sgRNA2 can aim at the same target gene or different target genes of the alfalfa. According to the invention, double targets are designed for a target gene according to the characteristics of an autotetraploid of alfalfa, a gene editing binary vector p6401-Target is constructed, and alfalfa is subjected to agrobacterium-mediated transformation to obtain a regenerated plant. According to the optimized genome editing system, the alfalfa gene editing efficiency is greatly improved, the plant gene editing efficiency can reach up to 100%, and the single target editing efficiency can reach up to 96.9%.

The invention relates to a plant expression vector for the specific expression of epatitis B surface antigen (HBsAg) in fruits. The vector adopts the promoter of agglutinin gene expressed specificallyin banana fruits instead of the constitutive 35S promoter on the plant expression vector pBI121, replaces the gus gene on the pBI121 with HBsAg gene and constructs recombinant plant expression vectorpCAMBIA2300HBS. Then agrobacterium infection method is used to transform the vector to transform the banana, PCR detection proves that exogenous gene is inserted in banana genome; and ELISA detectionproves that the promoter of banana lectin gene drives the specific expression of HBsAg gene in fruits. The acquisition of the vector can provide a favorable tool for the specific expression of HBsAggene in fruits and specially lay the foundation of using the vector to prepare hepatitis B edible vaccine. Therefore, the vector has important application value in the future transgenic research.

Provided are systems, constructs, genetically modified organisms, and methods for creating transgenic rodent research and commercial models of human physiology, disease, syndromes, and disorders. Provided are genetically modified rodents encoding for an sgRNA useful in a Cas9-mediated split gene-drive system for optimization of the gene drive system in rodents.

An insect gene drive system for biocontrol of a population of an insect is provided. The gene drive system includes: a) a first DNA sequence encoding a toxin under the control of a maternal germline-specific promoter active in the insect, with the first DNA sequence being linked to b) a second DNA sequence encoding an antidote under the control of an early embryo-specific promoter active in the insect. The toxin is expressed in maternal germline cells of the insect and results in maternal-effect lethality in the insect, and the antidote is expressed in embryos of the insect and counters the maternal-effect lethality. In some embodiments, the insect is Drosophila suzrukii.

A seed-specific expression vector and its construction methods and applications are disclosed. A fusion protein expression cassette consisting of Arachis hypogaea oleosin gene-apolipopoprotein A-IMilano (A-IM) gene driven by Brassica napus oleosin gene promoter is inserted between the HindIII and SacI sites of a plant binary expression vector pBI121, obtaining the plant expression vector pBINOA of the invention. In addition, a method for producing apolipoprotein A-IMilano is provided, in which the expression vector is used to transform oil sunflower which is used as a plant bioreactor. The method can not only improve the yield of apolipoprotein A-IMilano, but also greatly reduce production costs, and is suitable for industrial production.

The invention discloses an abiotic stress inducible promoter and an application thereof. The invention specifically discloses a DNA molecule. The DNA molecule is an inducible promoter PHSPIn9 and an inducible promoter PHSPDel. The promoter is derived from a GhHSP70 gene dominant haplotype promoter in upland cotton, the expression quantity of a GUS gene driven by the promoter under the abiotic stress is detected, and the result shows that both the PHSPIn9 promoter and the PHSPDel promoter can enhance the expression level of the GUS gene under the abiotic stress condition. The relative expression level of the GUS gene driven by the PHSPIn9 promoter is remarkably higher than that of the PHSPDel and CaMV35S. The developed abiotic stress inducible promoter provides an excellent tool for directional expression of genes for genetic engineering breeding of plants for coping with abiotic stress such as high temperature and drought, and has important significance for research and production practice of a plant genetic engineering technology.

The invention belongs to the technical field of plant molecular biology, and in particular discloses a shepherd's purse peroxidase gene promoter and an application thereof in improving plant cold tolerance. The nucleotide sequence of the shepherd's purse peroxidase gene promoter is that of the 1st-1021st bases in SEQ ID No.1. A gene driven by the promoter can code a peroxidase gene which is inducible by low temperature and is capable of regulating reactive oxygen in plant cells. Under an induction condition of 4 DEG C, the expression amount of the promoter is up-regulated, and meanwhile, the expression amount of the promoter is also up-regulated in leaves and stems. An inducible plant expression vector, which is constructed by virtue of promoter fragments, can start the expression of a target gene in the case that plants are under low-temperature stress. The invention also provides an application of the promoter in breeding cold-tolerance plants, so that the cold tolerance of crop varieties can be improved.

The invention discloses the application of the CRISPR / Cas9 genome editing system in alfalfa gene editing. The CRISPR / Cas9 genome editing system includes an expression vector, and the expression vector may include a Cas9 expression cassette, an sgRNA1 expression cassette, and an sgRNA2 expression cassette. The Cas9 expression cassette expresses Cas9. The sgRNA1 expression cassette expresses the sgRNA named sgRNA1, and the sgRNA2 expression cassette expresses the sgRNA named sgRNA2; the sgRNA1 expression cassette contains a promoter named MtU6-6promoter and the sgRNA1 gene driven by the MtU6-6promoter, The sgRNA2 expression cassette contains a promoter named MtU6‑5promoter and the sgRNA2 gene driven by the MtU6‑5promoter; the sgRNA1 and the sgRNA2 can be aimed at the same target gene or different target genes of Medicago sativa. According to the characteristics of alfalfa autotetraploid, the present invention designs dual targets for target genes, constructs a gene editing binary vector p6401-Target, and transforms alfalfa through Agrobacterium-mediated transformation to obtain regenerated plants. This optimized genome editing system has greatly improved the gene editing efficiency of alfalfa. The gene editing efficiency of plants can be as high as 100%, and the editing efficiency of a single target can be as high as 96.9%.

The invention relates to cloning and application of a peanut constitutive promoter, belonging to the technical field of biology. A constitutive promoter of peanut AhLEC1 gene is cloned from the peanut, and the gene sequence is disclosed as SEQ ID NO:1. The GUS histochemical stain proves that the drive gene of the promoter forms constitutive efficient expression in roots, stems, leaves, flowers, embryos and other organs of plants. On the basis of the function of the promoter, a plant expression vector of GUS report gene driven by the promoter is constructed and used for transforming Arabidopsis thaliana, which indicates that the promoter has similar drive effects on the GUS gene to CaMV35S. The promoter has moderate segment size, is convenient for construction, and is suitable for application in laboratories and industry.

The invention discloses a vector, a transformant and an application thereof comprising a vascular bundle tissue-specific expression promoter. The promoter is obtained from the sugar beet heart genome, and its nucleotide sequence is shown in SEQ ID: No 1. The promoter can significantly promote the expression of the reporter gene GUS in the vascular tissue of Arabidopsis, and it has a specific expression effect in the vascular tissue of the plant, so that the disease resistance gene driven by the promoter can be expressed in the vascular tissue of the plant. It can be expressed in the medium to resist Fusarium wilt and other pathogens, which provides a new way for plant disease-resistant breeding.

The present invention provides novel functional assay for 5-HT2A, histamine H1 or adrenergic alpha 1b receptors, by measuring intracellular cyclic adenosine monophosphate (cAMP) levels utilizing reporter gene driven cell based assay. The novel assay provides both binding affinity as well as mode of action of compounds in a single set. The novel assay of the invention is useful in identification of compounds acting through 5-HT2A, histamine H1 or adrenergic alpha 1b receptors. Furthermore, the assay offers utility in categorizing compounds in to agonist, partial agonist, inverse agonist and antagonist classes. The novel assay can be scaled up to any high throughput format.

The invention provides a method for constructing a B cell deficient mouse model and its application in B cell deficient drugs. This method inserts the diphtheria toxin receptor and green fluorescent protein into the Cd19 molecular gene expression cassette, and constructs a Cd19-IRES-DTR-EGFP (abbreviation: Cd19-DTR) gene-modified mouse model, which can realize intracellular The source Cd19 gene drives the expression of diphtheria toxin receptor and green fluorescent protein. Using this model, B cells can be killed by administering diphtheria toxin to mice at a specific time point or period of time, resulting in B cell defects, and the role of B cells in specific physiological and pathological processes can be studied.