632 results about "Intron" patented technology

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to the testis size character of Hu sheep as well as a screening method and application of the SNP molecular marker, and belongs to the fields of genetic breeding and molecular biology. The marker is located in an intron region of a Hu sheep chromosome 3 LRRIQ1 gene and is remarkably related to the Hu sheep testis size character, and the accuracy and reliability of the SNP molecular marker are ensured through a scientific and rigorous screening method; according to the marker, Hu sheep individuals related to testis size characters can be rapidly and accurately screened out, the breeding efficiency and accuracy are improved, the breeding process is accelerated, the breeding cost is reduced, and the economic benefits are improved; the genetic diversity is protected, the accuracy of seminal fluid quality evaluation is improved, a powerful tool is provided for Hu sheep breeding work, and important basic data is also provided for researching a genetic mechanism of testis development and functions; the method can also be used as an auxiliary diagnosis tool to assist in identifying potential genetic defects or disease risks and promote progress and development of the breeding technology.

The invention discloses an STON2 gene intron SNP (Single Nucleotide Polymorphism) molecular marker related to egg laying traits of chickens and application of the STON2 gene intron SNP molecular marker, and belongs to the technical field of molecular genetics. The research finds that the STON2 gene may participate in the related process of poultry sexual precocity and egg laying, so that the reproductive performance and the egg laying character of the chicken are influenced, and four SNP loci remarkably related to the egg laying character of the chicken are found in the fifth intron region and the sixth intron region of the STON2 gene for the first time. Based on the SNP site, the invention further develops an SNP molecular marker related to the egg laying traits of the chickens, and the SNP molecular marker can be used for auxiliary breeding of high-yield laying hens.

According to the invention, a structural domain reaction substrate sequence consisting of Exon1 (E1), P1 and Exon2 (E2) sequences of ribozyme is mutated, and E1, P1 and E2 sequences are mutated under the condition of maintaining the structural stability, so that the Azoarcus group I intron ribozyme still has enzyme activity and can maintain the capability of forming circular RNA (Ribose Nucleic Acid). The invention discloses a flexible vector construction method for preparing circular RNA (Ribonucleic Acid) without limitation of a substrate sequence, which comprises the following steps: determining a target to-be-cyclized site sequence NNUNNNN, and segmenting the target to-be-cyclized site sequence NNUNNNN into E1: NNU and E2: NNNN; with E1 and E2 sequences as references, designing IGS sequences to respectively form complementary pairing with E1 and E2; and 5'and 3 'homologous arms, IRES, CDS and other elements are respectively added. The method provided by the invention can be used for preparing the circular RNA for any target sequence, has no residual sequence, and has relatively high cyclization efficiency. The FlexCirc cyclization system designed on the basis of Azoarcus group I intron ribozyme can form the circular RNA, the cyclization substrate sequence has the characteristic of flexible design, and the cyclization efficiency can realize a relatively high cyclization proportion.

The invention provides an hpRNA interference vector for laver and application of the hpRNA interference vector, and belongs to the technical field of genetic engineering and algae biology. According to the hpRNA interference vector, a binary vector pBI121 is used as a skeleton vector, a laver endogenous Actin promoter and a multiple cloning site segment are inserted, the multiple cloning sites allow target gene segments to be cloned in a forward and reverse repetition mode, the target gene segments are spaced by introns, and therefore an efficient hpRNA structure is formed after transcription, and the efficient hpRNA interference vector is obtained. The gene can be directly used for silencing important genes related to growth and development, metabolic regulation, stress response and the like in the laver. The invention further provides a construction method of the hpRNA interference vector and application of the hpRNA interference vector in silencing laver genes and provides a method for silencing laver gene expression, the gene silencing effect is remarkable, experimental results can be repeated, and a powerful tool is provided for molecular breeding and genetic improvement of laver.

The present invention relates to a viral gene therapy vector for removing hepatitis B viruses, a preparation method therefor, and the use thereof, aiming to solve the problem in which existing viral gene therapy vectors can only inhibit the replication of hepatitis B viruses, but cannot remove virus cccDNA. The vector comprises a promoter, an intron, an enhancer, a nuclear localization signal (NLS) functional domain encoding gene of PGLYRP2 protein, an HBV DNA binding domain encoding gene of the PGLYRP2 protein, a secretory signal peptide functional domain encoding gene of the PGLYRP2 protein, an HBV nucleocapsid-binding functional domain encoding gene of the PGLYRP2 protein, and an IRES fragment. The viral gene therapy vector can remarkably promote the removal of HBVs in the liver of mice and is used for treatment of hepatitis B virus infection.

The invention discloses an SNP molecular marker related to dry and wet environment adaptability of sheep and application of the SNP molecular marker, and belongs to the field of molecular biology. The SNP molecular marker related to the humid environment adaptability of the sheep is located in an intron region of a BMPR1B gene, the gene mutation of the site significantly affects the sheep, and when the genotype of the site is AA, the sheep is the sheep with dry environment adaptability; when the genotype of the locus is GG, the locus is a sheep adaptive to a humid environment, so that a new molecular breeding marker is provided for screening or introducing the sheep adaptive to the humid environment.

The invention discloses a method, a system and equipment for detecting internal tandem repeat and a storage medium, and the key points of the technical scheme are as follows: obtaining a first reference sequence according to at least one target exon sequence corresponding to a protooncogene, and obtaining a second reference sequence according to at least one target intron sequence corresponding to the protooncogene; comparing the sequencing data of the to-be-detected sample to the second reference sequence to obtain a first comparison result, extracting an uncompared sequence from the sequencing data according to the first comparison result, and comparing the uncompared sequence to the first reference sequence to obtain a second comparison result; and determining a first detection result according to the second comparison result and a first reference sequence, and performing false positive filtering on the first detection result to obtain a second detection result. According to the invention, false positive can be reduced so as to ensure the accuracy and reliability of subsequent analysis.

The present invention relates to an exogenous target gene expression cassette for delivering an exogenous target gene to the retina, in particular AIPL1 to retinal pigment epithelial cells and photoreceptor cells, comprising an IRBP enhancer sequence, a rhodopsin kinase (RK) promoter sequence and a CAG intron sequence, which are operatively linked, and an exogenous target gene. The present invention also relates to a recombinant adeno-associated viral vector comprising a viral capsid comprising a capsid protein or a capsid protein variant and a viral vector genome comprising an expression cassette encoding for specifically expressing an exogenous target gene in retinal pigment epithelial cells and photoreceptor cells. The recombinant adeno-associated virus vector can be used for relieving or treating retinal degenerative eye diseases by intravitreal administration or subretinal administration.

The invention provides an application of a genetic marker associated with a chest width character in an RNASEH2B gene in chicken genetic breeding, and belongs to the technical field of chicken genetic breeding and biology.The genetic marker associated with the chest width character in the RNASEH2B gene comprises CBmark er1, the CBmark er1 corresponds to the 169817100th site of the physical position of a first chromosome of chicken reference genome bGa < 1 >. Mat.broi ler.GRCg7b version sequence information published in NCBI, and the CBmark er1 corresponds to the 169817100th site of the physical position of a second chromosome of the chicken reference genome bGa < 1 >. Mat.broi ler.GRCg7b version sequence information. The nucleotide sequence belongs to the ninth intron sequence of the gene RNASEH2B, and the basic group in the ninth intron sequence is G or A; the CBmarker1 contributes to heredity improvement of the chest breadth, and when the CBmarker1 is applied to genetic breeding of chickens, improvement of the chest breadth is facilitated, and a high-quality laying hen variety is obtained.

The invention relates to a high-throughput cis-acting element screening carrier and a screening method. Specifically, the invention provides a plasmid vector system containing bar codes, each bar code in the system is in one-to-one correspondence with a candidate cis-acting element, and the activation multiple of the candidate cis-acting element can be obtained by measuring the abundance of the bar codes; in order to eliminate the influence of a bar code on the vector on a detection result, an exogenous intron which can be cut off during transcription is inserted into a coding region in the vector and is used for distinguishing vector DNA and RNA obtained by transcription. The screening method of the biological cis-acting element has high efficiency, wide applicability and high throughput, and has outstanding application value in biological research and breeding.

The invention relates to an SNP molecular marker related to chicken first laying day age and egg yield characters and application thereof, and belongs to the technical field of genetics, the SNP molecular marker is derived from an FST gene, the nucleotide sequence is as shown in SEQ ID NO.1, and the sequence as shown in SEQ ID NO.1 mutates into G (namely 1493 (Agt) on Enmbl FST sequence ENSGALG00010012441) from the 1493th basic group A at the 5'end; g)). According to the invention, the fact that the chicken FST gene Intron 1 has one mutation site is found for the first time, that is, the 500-day-old egg laying number of A1493G and GG genotypes is higher than that of other genotypes, and the GG genotype individuals are selected for breeding reservation, so that the overall production performance of chicken flocks can be improved. Therefore, by detecting the molecular marker associated with the egg laying traits, the method is simple, convenient and rapid, breeding of high-yield laying hen varieties is facilitated, and the breeding process is accelerated.

The invention provides gRNA (guide Ribonucleic Acid) targeting an AGT (Absorption Growth Tube) gene and application thereof, in particular, the invention provides gRNA targeting an AGT gene exon-intron splicing junction region, the invention firstly discovers that the gRNA designed by targeting the AGT gene splicing junction region (such as an intron 1 receptor, an intron 2 donor / receptor and an intron 4 donor / receptor) can effectively inhibit the expression of the AGT gene, therefore, the gRNA can be used for inhibiting the expression of the AGT gene. According to the gRNA designed by the invention, the gene editing efficiency can be remarkably improved; and / or treating a disease.

Use of PGLYRP2 gene and a related drug thereof. The present invention relates to the field of biopharmaceuticals, and in particular, to use of the PGLYRP2 gene and the related drug thereof. The present invention aims to solve the problem that existing drugs can only inhibit the replication of hepatitis B virus and cannot eliminate viral cccDNA. The PGLYRP2 gene is used for preparing a drug for treating a hepatitis B virus infection. The PGLYRP2 gene is used for preparing a drug for eliminating hepatitis B virus cccDNA. Also provided is a gene therapy vector for eliminating hepatitis B virus. The gene therapy vector comprises a promoter, an intron, an enhancer, and a PGLYRP2 gene encoding region. The HBV virus inhibition function and the cccDNA-eliminating effect of the PGLYRP2 protein are defined, so that the elimination of HBV in liver cells and a mouse liver can be significantly promoted.

The invention belongs to the technical field of molecular biology, and discloses an RNA cyclization composition for improving cyclization efficiency, and a preparation method and application of circRNA. The composition for RNA cyclization comprises a precursor fragment and a cyclization induction fragment, the cyclization induction fragment comprises an affinity arm I and an affinity arm II which are complementarily paired with two ends of a to-be-cyclized sequence respectively, and the space distance of 3 '-intron and 5'-intron is effectively shortened through the bridging effect of the cyclization induction fragment, so that the cyclization effect of the RNA is improved, and the cyclization effect of the RNA is improved. According to the present invention, the two transesterification reactions are introduced, such that the active sites for catalyzing the two transesterification reactions are close to each other, the precursor fragment has excellent in-vitro cyclization efficiency, and the circRNA obtained through cyclization has characteristics of extremely low exogenous nucleotide residue and high biological safety.

The invention discloses a molecular marker combination related to a cockscomb height character and a detection reagent and application thereof, and belongs to the technical field of molecular genetics. According to the molecular marker combination and the detection reagent thereof, ten SNP loci remarkably associated with the chicken first-laying crown height and the 130-day-old crown height are identified in the intron of the CTCF gene for the first time, based on the SNP loci, the molecular marker combination related to the cockscomb height character and the detection reagent thereof are designed, and the molecular marker combination and the detection reagent thereof can be used for early screening of laying hens with high cockscomb height and can be used for early screening of laying hens with high cockscomb height. The method is of great significance to breeding and breeding of precocious high-quality laying hens.

The disclosure relates to compositions, systems, and methods related to repair RNA sequences, as well as related systems for trans-splicing a target nucleic acid using a repair RNA sequence. Various compositions and systems are described herein, including compositions comprising a repair RNA sequence comprising one or more exons and / or introns, a splice donor and / or splice acceptor, wherein the repRNA is suitable for trans-splicing. The compositions, systems, and methods described herein are suitable for targeting one or more Usher syndrome-associated genes, and treating Usher syndrome.

The present invention relates to the field of molecular biology, and further relates to a specific marker, primer pair, detection method and application of DNA insertion variation in the intron region of ihrp gene of spotted rockfish. The specific marker of insertion variation in the intron region of ihrp gene of spotted rockfish is a DNA fragment of non-insertion and insertion specific marker in the intron region of ihrp gene of spotted rockfish, and the nucleotide sequence of the DNA fragment is shown in SEQ ID NO:1 and SEQ ID NO:2. The present invention utilizes a pair of primers to amplify bands of 546bp and 419bp in individuals with DNA insertion variation, and only amplifies a band of 419bp in individuals without insertion variation. The detection method of the present invention significantly shortens the identification time, improves the detection efficiency, and has important application value in sex identification, preparation of Kaohsiung seedlings and family selection of spotted rockfish.

The invention discloses an Indel marker related to a corn kernel protein content character and application of the Indel marker, and belongs to the technical field of plant molecular genetics and crop molecular design breeding. The Indel marker is located on a gdh2 gene of a chromosome 10 of a corn genome, and insertion or deletion of a nucleotide sequence as shown in SEQ ID NO.5 occurs in an intron region between a second exon and a third exon. By applying the molecular marker-assisted selection technology, high-protein new germplasms S3317 (the crude protein content is 17.0%) and JS231148 (the crude protein content is 15.6%) have been successfully cultivated, the increase amplitudes of the high-protein new germplasms and JS231148 are both more than 15% compared with those of the conventional control, and the lysine content reaches 0.41% and reaches the national high-quality protein corn standard.

The invention provides a DGAT1 gene SNPs marker for detecting sheep milk production traits and application thereof.Nucleotide sequence variation sites of the DGAT1 gene are detected with the first-filial generation of Dongfudrisheng sheep male * Hu sheep female as an object, a general linear mixing effect model is constructed, the correlation between nucleotide sequence variation and sheep milk production traits is researched, and the DGAT1 gene SNPs marker for detecting the sheep milk production traits is obtained. And digging a molecular genetic marker for regulating and controlling the lactation character of the sheep. Three SNPs (single nucleotide polymorphisms) sites are found in total; wherein two variation sites in the first intron are named as c.191 + 411 Cgt, and the second intron is named as c.191 + 411 Cgt; t (SNP1) and c.192-440 Cgt, and c. 292-440 Cgt; t (SNP2) and a variation site in the 17th exon are named as c.1461 Cgt; t (SNP3). For the SNP1 site, the average daily milk yield of the CT genotype ewe is increased by 8.2% and 17.04% compared with that of the CC genotype ewe and that of the TT genotype ewe respectively; for the SNP3 site, the milk fat percentage of the CC genotype ewe is improved by 0.314% compared with that of the CT genotype ewe. A new molecular marker can be provided for selection and improvement of lactation traits of the milk sheep, and the molecular marker has important application prospects in sheep screening or breeding.

The invention belongs to the technical field of gene detection, and particularly relates to application of a CFTR gene mutation combination, an amplification reagent and a product in preparation of a CF risk assessment product. The CFTR gene mutation combination consists of an intron region mutation combination and an exon region mutation combination, based on the CFTR gene mutation combination, the invention further develops an amplification reagent and a CFTR gene mutation detection product, the amplification reagent and the CFTR gene mutation detection product comprise a primer combination for multiplex PCR amplification of the CFTR gene mutation combination, and the primer combination has the characteristics of high accuracy, high specificity and high sensitivity, can accurately detect related gene mutation, and provides a reliable basis for risk assessment and diagnosis of CF. When the amplification reagent and the CFTR gene mutation detection product are combined with sweat chlorine detection for use, the diagnosis rate of CF can be remarkably improved, and the amplification reagent has important application value in clinical diagnosis of CF and is expected to provide more powerful support for early diagnosis and treatment of CF patients.

Colorectal cancer (CRC) has not benefited from innovative immunotherapies, mainly because of the lack of actionable immune targets. Novel tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs) expressed by CRC cells are described herein. Most of the TSAs described herein derives from aberrantly expressed unmutated genomic sequences, such as intronic and intergenic sequences, which are not expressed in normal tissues. Nucleic acids, compositions, cells, antibodies and vaccines derived from these TSAs are described. The use of the TSAs, nucleic acids, compositions, antibodies, cells and vaccines for the treatment of CRC is also described.

The present invention provides nucleic acids and methods for making circular RNAs (circRNAs), compositions comprising circular RNAs, and methods of use thereof. The present invention uses optimized self-splicing T4 bacteriophage td gene (T4td) intron sequences to produce circular RNAs. The optimized T4td intron sequences described herein mediate efficient circularization of a linear RNA sequence.

The present application provides methods for producing circular RNAs (circRNAs) from a DNA construct encoding a linear RNA precursor, wherein the linear RNA precursor comprises from the 5′-end to the 3′ end: a 3′ catalytic Group I intron fragment, a 3′ exon sequence, an effector RNA sequence, a 5′ exon sequence, and a 5′ catalytic Group I intron fragment, wherein the method comprises an in vitro single-pot reaction. In some embodiments, the single-pot reaction does not comprise supplementing the reagent composition with GTP, a divalent metal ion such as Mg2+, or DNase I prior to circularization of a linear RNA precursor.

The invention discloses an NDRV [sigma] C circular RNA vaccine and application thereof, and belongs to the technical field of vaccines.Duck hepatitis A virus type 1 (DHAV-1) UTR and T4 bacteriophage I type intron self-splicing cyclization systems are adopted to construct circular RNA (CirRNA-[sigma] C) for expressing NDRV [sigma] C protein, and a vaccine preparation is prepared through a chitosan-polyethyleneimine (CS-PEI) nano delivery system. The feasibility of a circular RNA vaccine system based on DHAV-1UTR in poultry vaccines is confirmed for the first time, the established nano delivery technology and circular RNA vaccine platform have the characteristics of high stability, good safety, strong immunogenicity and the like, and an important technical path is provided for development of novel vaccines of NDRV and other viral epidemic diseases.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to the tolerance character of white spot syndrome virus (WSSV) of procambarus clarkia, the molecular marker is located in the third intron region of a TRIM gene, is specifically located at 585bp of a DNA (Deoxyribose Nucleic Acid) sequence as shown in SEQ ID NO.1, and is Tgt; the T / C heterozygous genotype is obviously related to the WSSV tolerance character of the procambarus clarkia. The invention provides a favorable marker for breeding of disease-resistant molecules of procambarus clarkii, and can accelerate the breeding process of disease-resistant varieties of procambarus clarkii.

The invention relates to the technical field of biology, in particular to a tilapia mossambica streptococcus agalactiae resistance related KASP molecular marker and application thereof. According to the invention, two SNP (Single Nucleotide Polymorphism) loci, namely SNP-1 and SNP-2, which are remarkably associated with resistance are screened based on an intron region of an EGFRX2 gene of tilapia mossambica, a typing method of the loci is developed through a KASP technology, and relevance between the genotype of the SNP-1 and a resistance phenotype is verified. Furthermore, streptococcus agalactiae resistance dominant diplotype individuals are screened in combination with double SNP loci, and the survival rate of a challenge experiment is remarkably improved. The detection kit provided by the invention can realize high-throughput and high-precision typing, is suitable for tilapia mossambica resistance genotype identification and molecular marker-assisted breeding, provides an efficient genetic improvement tool for tilapia mossambica disease-resistant breeding, and is beneficial to reduction of breeding disease loss and reduction of antibiotic dependence.

Compositions and methods thereof for increasing endogenous FAN1 expression in a target cell are provided. Preferably, compositions include one or more splice switching antisense oligonucleotides targeting FAN1 exon 10 – intron 10 – exon 11 junction. The compositions are effective in increasing levels of functional FAN1 mRNA transcripts encoding FAN1 derived from the pre-mRNA in a target cell. The compositions and methods are particularly suited for treating, ameliorating, or delaying the onset of a trinucleotide repeat expansion disorder in a subject in need thereof.

The present invention provides an agent capable of modulating variable splicing in the intron of the SLC20A2 gene, and its use for modulating the expression of functional PiT2 protein to prevent and / or treat disease. The invention further provides a brain calcification model based on variable splicing in the intron of the human SLC20A2 gene, a preparation method of the brain calcification model, and application of the model to indication and analysis of the incidence stage of brain calcification, or to a model for researching phosphorus homeostasis imbalance in brain or a target of brain calcification, or to screening of drugs.

DNA molecules for making a circular RNA are provided. The DNA molecule may include elements operably connected and arranged, from a 5′ to 3′ direction, in the following order: (a) an intron fragment that includes a full-length intron; (b) an E2 fragment which includes a downstream exon of the full-length intron; and (c) an E1 fragment which includes an upstream exon of the full-length intron. 3′ end of the E1 fragment is configured to produce a hydroxyl group in an in vitro transcription reaction; the hydroxyl group is capable of initiating splicing in a one-step transesterification reaction at a splice site between RNA fragments transcribed from the intron fragment and the E2 fragment in a linear RNA that is produced from the DNA molecule in the in vitro transcription reaction, such that the linear RNA is configured to self-circularize to produce the circular RNA.

Disclosed herein are nucleic acid compositions and methods of use. The nucleic acid compositions may have a therapeutic nucleic acid sequence operably linked to a nuclear targeting sequence that increases expression of the therapeutic nucleic acid in a cell by at least 1.25 fold; at least 80% sequence identity to SEQ ID NO: 6; or a first regulatory element comprising a promoter sequence operably linked to a hemoglobin subunit gamma intron (hBGi) sequence, and a second regulatory element comprising a woodchuck hepatitis posttranscriptional regulatory element (WPRE) sequence. The nucleic acid compositions with these features may enhance therapeutic nucleic acid transfection and expression. Also disclosed herein are transgenes optimized for gene therapy applications, including novel FVIII transgene sequences, in which the B domain may be non-naturally occurring the A1 and / or A3 domain may include at least one amino acid substitution.