20 results about "Nonsense mutation" patented technology

The invention belongs to the technical field of biology, and particularly relates to ANK1 gene nonsense mutation and application. The invention firstly provides an ANK1 gene non-sense mutation c.2230 Cgt and an ANK1 gene non-sense mutation c.2230 Cgt. The invention relates to the field of genetic engineering, and in particular relates to nonsense mutation T (p.Q744X), the nonsense mutation significantly reduces the expression of ankyrin, and further causes phenotypic changes such as abnormal cell morphology and increased osmotic fragility after K562 erythroid differentiation, which indicates that the K562 is related to the pathological process of hereditary polycythemia spheroides (HS), and indicates that the mutation has pathogenicity; secondly, a treatment evaluation system taking drug-induced translation readthrough and adenine base editing as a core is constructed around the mutation, a new strategy and a technical platform are provided for precise molecular treatment of the ANK1 nonsense mutant HS, and important clinical transformation prospects and application values are achieved.

The invention discloses a DNA shuffling method based on flanking sequence segmented amplification and application of the DNA shuffling method in screening herbicide-resistant hybrid enzymes, on the basis of a single-stranded DNA shuffling technology, the success rate of single-stranded preparation is increased by introducing flanking sequences and segmented amplification recombinant products into single-stranded amplification primers, and the yield of herbicide-resistant hybrid enzymes is increased. Meanwhile, the proportion of nonsense mutation in the mutation library is reduced. The method is expected to provide a new method for establishing a protein qualitative evolution mutation library. The method is used for shuffling the rice and corn hppd genes and screening to obtain the hybrid enzyme of which the resistance is stronger than that of parent rice and corn HPPD under the treatment of 300 mu M mesotrione.

The invention provides a PMFBP1 mutant gene related to azoospermia syndrome and an application of the PMFBP1 mutant gene. The mutant gene is a PMFBP1 gene c.2641Cgt; the protein p.Arg881Ter is subjected to non-sense mutation, so that the encoded protein p.Arg881Ter is truncated, and the function of a sperm head-tail connection device is damaged. The invention also provides a specific Sanger sequencing detection method and a kit of the mutation, which can be used for rapid molecular diagnosis, genetic counseling and assisted reproduction guidance of the disease, and provides a basis for precise medical treatment.

The invention discloses an editing system for specifically repairing a pathogenic mutant gene by combining Sup-tRNA with gsnoRNA. According to the method, the Sup-tRNA and the gsnoRNA are combined to repair the pathogenic nonsense mutation for the first time, so that the mutation read-through efficiency is improved, and off-target read-through is avoided. In addition, the invention also provides a recombinant vector and a cell comprising the editing system, and provides a pharmaceutical composition. The invention provides a new strategy for treating pathogenic nonsense mutation related diseases, and has a wide application prospect clinically.

The present disclosure relates to compositions and methods for treating genetic disorders caused by nonsense mutations using anticodon-engineered transfer RNA (ACE-tRNA) constructs. These DNA-based ACE-tRNA constructs are designed to suppress premature termination codons (PTCs) and restore the expression of full-length, functional proteins. The disclosure further provides formulations of ACE-tRNA constructs with poly(amine-co-ester) (PACE) polymeric nanoparticles to improve stability, protect nucleic acids, and enhance delivery to airway epithelial cells. Also described are functionalized ACE-tRNA Picovectors (sLPVs) incorporating targeting elements such as nuclear localization signals (NLSs), nucleolar localization sequences (NoLSs), and DNA nuclear targeting sequences (DTSs) to improve nuclear import and localization.

This invention relates to a method for constructing a mouse model of autosomal dominant polycystic kidney disease and its applications, particularly to a method for preparing a non-human animal model carrying a nonsense mutation in the Pkd1 gene c.616_618GAG>TAG based on CRISPR / Cas9 gene editing technology. The method involves co-injecting mouse zygotes with gRNA targeting exon 5 of the Pkd1 gene, homologous recombinant donor oligonucleotides containing the c.616_618GAG>TAG mutation, and Cas9 nuclease. Mutant mice are obtained via embryo transfer. The mutation site is verified by PCR combined with sequencing, and a stable mutant line is established through breeding. This invention also includes the application of this model animal as a research tool for autosomal dominant polycystic kidney disease, particularly in the areas of pathogenesis analysis, drug screening, and therapeutic target validation. This model can stably simulate the typical clinical manifestations and pathological features of human autosomal dominant polycystic kidney disease, providing an important tool for basic research and translational medicine of this disease.

The application belongs to the technical field of molecular markers, and provides a Hu sheep 5K low-density SNP chip and application thereof.The chip comprises 5000 SNP molecular markers, and the SNP molecular markers are shown in Table 1.Based on resequencing data of Hu sheep, the application screens SNP markers with rich polymorphic information content of Hu sheep breeds, integrates functional variation sites such as nonsense mutations and splicing mutations, and customizes the 5K low-density chip of Hu sheep, which can be used for Hu sheep kinship identification and functional harmful mutation screening.

This invention relates to a gene encoding coagulation factor IX. F9 This study investigates animal models of nonsense mutations, methods for screening hemophilia drugs using these models, and the application of the screened stRNAs in the preparation of hemophilia drugs. Based on an analysis of the probability of nonsense mutations in coagulation factor IX in hemophilia patients, the study verifies that stRNAs can read the coagulation factor IX encoding gene. F9 The function and efficiency of the premature termination codon (PTC) were investigated. Seven Arg-stRNAs were designed to target the coagulation factor IX R75* mutation, and readthrough efficiency was tested at the cellular level. A mouse model of the coagulation factor IX R75* mutation was constructed, and Arg-stRNA was delivered via AAV, restoring the expression of endogenous coagulation factor IX and reducing clotting time, demonstrating the effectiveness of stRNA in treating hemophilia B.

The invention discloses application of single-base nonsense mutation of a sheep FUT2 gene in diarrhea resistance, and belongs to the technical field of diarrhea-resistant breeding of sheep. The invention provides a single base mutation related to diarrhea resistance of sheep, which is located at the 55082566th nucleotide from the 5'end of chromosome 14 of a sheep genome AR-UIRamv2.0 version, and when the site is T, the sheep shows the ability to resist infectious diarrhea; the provided single base mutation is linked with sheep anti-diarrhea, can be used as a molecular marker, is used for breeding infectious diarrhea resistant sheep varieties and / or screening natural anti-diarrhea individuals, is extremely high in accuracy, strong in universality and simple and convenient to operate, can realize fundamental transformation from post phenotype selection to beforehand genotype selection, and has a wide application prospect. The method has important significance in simplifying the breeding process, shortening the breeding period and workload and improving the breeding efficiency.

The invention relates to a construction method and application of an autosomal dominant polycystic kidney disease mouse model, in particular to preparation of a Pkd1 gene carrying c.616618GAGgt gene based on a CRISPR / Cas9 gene editing technology. The invention relates to a method for a non-human animal model of TAG nonsense mutation. The construction method comprises the following steps: co-injecting gRNA (guide Ribonucleic Acid), containing c.616618GAGgt, of a No. 5 exon of a targeted Pkd1 gene into a mouse fertilized egg; carrying out embryo transplantation on TAG mutated homologous recombination donor oligonucleotide and Cas9 nuclease to obtain a mutated mouse; a mutation site is verified by combining PCR (Polymerase Chain Reaction) with sequencing, and then a stably inherited mutation line is established by breeding. The invention also comprises application of the model animal as an autosomal dominant polycystic kidney disease research tool, especially application in the fields of pathogenesis analysis, drug screening and therapeutic target verification. The model can stably simulate typical clinical manifestation and pathological characteristics of human autosomal dominant polycystic kidney disease, and provides an important tool for fundamental research and transformation medicine of the disease.

The present disclosure relates to compositions and methods for treating genetic disorders caused by nonsense mutations using anticodon-engineered transfer RNA (ACE-tRNA) constructs. These DNA-based ACE-tRNA constructs are designed to suppress premature termination codons (PTCs) and restore the expression of full-length, functional proteins. The disclosure further provides formulations of ACE-tRNA constructs with poly(amine-co-ester) (PACE) polymeric nanoparticles to improve stability, protect nucleic acids, and enhance delivery to airway epithelial cells. Also described are functionalized ACE-tRNA Picovectors (sLPVs) incorporating targeting elements such as nuclear localization signals (NLSs), nucleolar localization sequences (NoLSs), and DNA nuclear targeting sequences (DTSs) to improve nuclear import and localization.

The invention belongs to the field of molecular biology, and provides a functional molecular marker of a major gene AhNAC2 for regulating and controlling the oil content of peanut seed kernels and application of the functional molecular marker. The invention discloses a major site qSOCA08-1 for regulating and controlling the oil content of peanut seed kernels, and the phenotypic variation of 11.41-20.97% is explained. The qSOCA08-1 is finely positioned in a 352.2 kb physical interval between a No.8 chromosome marker Tif2. A08.38582892 and a No.8 chromosome marker Tif2. A08.38935104, and a candidate gene of the qSOCA08-1 is confirmed to be AhNAC2. The invention also discloses a method for identifying the qSOCA08-1. The 285th basic group of the coding sequence of the AhNAC2 shows C-A nonsense mutation between parents Jihuansweet No.1 and W191, and the early termination can cause the function loss of the AhNAC2. A competitive allele specific PCR molecular marker is developed on the basis of a C-A mutation SNP site, the association between the molecular marker and the oil content of the seed kernel is successfully verified in a peanut germplasm resource material and a Yuhua 15 * ST001 recombinant inbred line population, and the important value of the marker in marker-assisted selection breeding of the high oil content of the seed kernel of the peanut is verified.

The invention discloses application of a PTCHD1 gene mutant and a detection reagent. Belongs to the technical field of gene mutation detection. According to the mutation, compared with a wild type PTCHD1 gene, cytosine C is replaced with adenine A at the 1644th nucleotide of a coding sequence, so that protein translation is terminated in advance at the 548th amino acid, and a truncated protein is formed. And the c.1644Cgt of the PTCHD1 gene is confirmed; a is pathogenic nonsense mutation causing AUTS4, a new insight is provided for the genetic basis of the disease, and the blank of the mutation spectrum of the disease is filled.

The invention belongs to the technical field of biological medicine and gene therapy, and particularly relates to a tRNA binding protein assisted termination codon readthrough system and application thereof. The tRNA binding protein assisted termination codon reading-through system disclosed by the invention comprises inhibitory tRNA and binding protein thereof, wherein the inhibitory tRNA contains 70 pairs of G3: U, can be recognized by an endogenous alanyl-tRNA synthetase (AlaRS) of a cell, and is loaded with alanine; the tRNA binding protein is specifically bound with any structure, except for a receptor arm, of the corresponding tRNA, and can be specifically bound with a tRNA binding structural domain of pyrrole lysyl-tRNA synthetase. According to the system, alanine can be introduced in the translation process of a target gene carrying nonsense mutation, so that full-length translation of protein is recovered. The system can be applied to functional recovery and treatment research of various genetic diseases caused by the early termination codon.

The invention discloses an SEC23B gene mutant capable of reducing the content of SEC23B protein and application of the SEC23B gene mutant. The nucleotide sequence of the SEC23B gene mutant capable of reducing the content of SEC23B protein is shown as SEQ ID NO.3. Compared with an SEC23B gene with the nucleotide sequence shown as SEQ ID NO.1, the SEC23B gene mutant capable of reducing the content of SEC23B protein has nonsense mutations (c.181Tgt; c and c.1832 Ggt; according to the SEC23B gene capable of reducing the SEC23B protein content, the SEC23B gene is (A), cysteine at the 61th site of an SEC23B gene coding protein sequence is changed into arginine (p.C61R), and arginine at the 611th site of the SEC23B gene coding protein sequence is changed into glutamine (p.R611Q) through mutation. According to the SEC23B gene capable of reducing the SEC23B protein content, an SEC23B gene mutant with the SEC23B protein content is provided for the first time through systematic research, and the SEC23B protein content is reduced by 50% compared with a control group.

Provided herein are methods and compositions for modulating CNOT3 recruitment in a cell ribosome. The methods and compositions can modulate a target mRNA stability and / or half-life. Also provided are pharmaceutical compositions for use in the treatment of an mRNA-decay associated disease, nonsense mutation associated disease, obesity, a mitochondrial- associated disorder, or a metabolic disorder.

The invention relates to the skipping of the CEP290 exon 36 in an individual suffering from a retinal dystrophy accounted for by a nonsense mutation or a premature termination codon generated by a frameshift mutation in exon 36 or an upstream exon, including the c.4723A>T, c.4771C>T, c.4714G>T, c.4786_4790del, c.4791_4794del, c.4732G>T, c.4625_4626insCATG (35), c.4792_4795del, c.4801C>T, c.4805C>T, or c.4811G>A mutations, to bypass protein truncation and lessen retinal damages. Here, studying fibroblasts from control individuals, and two patients carrying the CEP290 c.4723A>T nonsense mutation, they show low levels of spontaneous skipping of exon 36 arising from both endogenous basal skipping and mutation-induced skipping. The minimally shortened and mutation-free CEP290 mRNA produced by skipping of exon 36 in the fibroblasts of the two patients is translated into a protein isoform that localizes at the centrosome and allows the formation of primary cilia, yet with elongated axonemes. Using an AON consisting of a sequence set forth as SEQ ID NO: 1, complementary to a nucleic acid sequence of CEP290 pre-mRNA, wherein said AON targeting an mRNA encoding the donor splice site (H36D) is capable to alter splicing by blocking the recognition of exon 36 and bypass protein truncation while maintaining the open reading frame, leading to the production of near full-length CEP290 protein, they were able to increase the abundance of the alternatively spliced mRNA and shortened protein and to reduce axonemal length in patient cells.

The invention discloses a construction method and application of an angel syndrome mouse model. The construction method comprises the following steps: preparing c.1435Agt carrying a maternal Ube3a gene on the basis of combination of a pilot editing technology and a tetraploid compensation technology; the invention relates to a non-human animal model of T nonsense mutation. The method comprises the following steps: co-delivering pegRNA (Ribonucleic Acid) and PEmax of a No.4 exon of a targeted Ube3a gene to a mouse embryonic stem cell, and introducing c.1435Agt at a target site; carrying out T nonsense mutation, and after tetraploid compensation technology treatment, transplanting the edited recombinant embryo to a pregnant female mouse to obtain an F0-generation mutant mouse; the accuracy of a mutation site is verified through combination of PCR amplification and gene sequencing, and a mutant strain with genetic stability is established through two generations of breeding and screening by using maternal genetic characteristics of the UBE3A gene. The invention further comprises a genotype identification method of the model animal, Ube3a gene mRNA expression quantity evaluation, behavioral evaluation and the like, and the Ube3a: c.1435 Agt is verified; the T mutant mouse model accords with the pathological characteristics of the angel syndrome, and can stably simulate the typical clinical manifestation and pathological characteristics of the angel syndrome.

The present disclosure relates generally to compositions and methods for treating, preventing, or slowing the rate of development of a disease or condition mediated by a nonsense mutation in the bone morphogenetic protein receptor type II (Bmpr2) in a subject in need thereof. The method entails administering to the subject a compound of the present disclosure, such as GJ103 and a salt thereof.

This disclosure provides Staphylococcus bacteriophages comprising a knockout and / or nonsense mutation in open reading frame 141 (ORF141), or a homologous position thereof. Also provided are compositions comprising the bacteriophage, methods of treating a bacterial infection, and methods of making a bacteria more susceptible to β-lactam antibiotic drug treatment.