342 results about "Intron" patented technology

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 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 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 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 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.

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 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.

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.

This vector for nucleic acid introduction contains a nucleotide sequence that encodes a Cas protein, a nucleotide sequence that encodes a guide RNA, and a nucleotide sequence of a nucleic acid to be introduced. The guide RNA targets an intron adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon. Also provided is a nucleic acid introduction method comprising a step for introducing a nucleic acid to be introduced through non-homologous end joining by targeting an intron that is adjacent to the 5' side, with reference to a sense strand, of an exon that includes a stop codon.

The present invention provides nucleic acids and methods for making circular RNAs (circRNAs), and circular RNAs, compositions and methods of use thereof. The present invention uses selected self-splicing intron sequences (e.g., intronic sequences derived from self-splicing Group I or Group II introns) to produce circular RNAs. These self-splicing intron sequences described herein mediate efficient circularization of a linear RNA sequence.

The present invention relates to chimeric promoters without CpG motifs. The chimeric promoter comprises a CMV enhancer, an IFN-gamma gene core promoter and an intron sequence, and a CG motif in the CMV enhancer, the IFN-gamma gene core promoter and the intron sequence is subjected to substitution mutation. The promoter can improve the expression efficiency of exogenous genes in activated immune cells.

The invention belongs to the field of biological medicine, and relates to a guide agRNA for regulating RNA splicing. The invention provides an application of a guide agRNA (Ribonucleic Acid) in preparation of a medicine for changing expression of a target protein by cells of a subject. The guide agRNA recruits ADAR protein to the 3 'splice site or pseudo 3' splice site to edit the A base of the 3 'splice site or pseudo 3' splice site, thereby splicing the whole exon located at the 3 'splice site or flanking the pseudo 3' splice site of the intron from the precursor mRNA, thereby changing the level of mRNA encoding the target protein, and altering the expression of the target protein in the cell. The target gene pre-mRNA selective splicing is regulated and controlled to influence the function or expression of the functional RNA and the target protein of the target gene, so that the purpose of treating diseases is achieved.

The present invention relates to Split-Intron Final Self-Inactivating (SIN) retroviral vectors which comprises a viral major splice donor (mSD) comprising mutations, a mouse mammary tumor vector long terminal repeat (MMTV-LTR) or a Type B leukemogenic virus long terminal repeat (TBLV-LTR), a eukaryotic splice acceptor (eSA), and a eukaryotic splice donor (eSD). Furthermore, the invention relation to the use and methods of uses of such vectors in gene therapy.

Provided is a method for producing a heterodimeric protein such as a bispecific antibody with which a heterodimeric protein composed of only domains having natural amino acid sequences can also be produced. A production method of the present invention is a method for producing a heterodimeric protein. The production method includes a production step of producing a heterodimeric protein by reacting a dimeric protein having a reaction tag with a modification protein for modifying the dimeric protein. The dimeric protein having a reaction tag includes a first monomeric protein and a second monomeric protein. The first monomeric protein includes a first reaction tag and a first dimer formation domain capable of forming a dimer in this order. The first reaction tag includes a binding tag and a first C intein. The second monomeric protein includes a second reaction tag and a second dimer formation domain capable of forming a dimer together with the first dimer formation domain in this order. The second reaction tag includes a binding partner capable of binding to the binding tag, and a second C intein. The modification protein includes a first modification protein and a second modification protein. The first modification protein includes a first N intein capable of reacting with the first C intein, and a first addition component to be added to the first monomeric protein. The second modification protein includes a second N intein capable of reacting with the second C intein, and a second addition component to be added to the second monomeric protein. The first addition component and the second addition component are different addition components. The first monomeric protein and the second monomeric protein form a dimer. In the production step, the first N intein of the first modification protein reacts with the first C intein of the first monomeric protein, and then the first addition component of the first modification protein is linked to the first monomeric protein, and the second N intein of the second modification protein reacts with the second C intein of the second monomeric protein, and then the second addition component of the second modification protein is linked to the second monomeric protein.

The invention discloses a CHRM3 gene InDel molecular marker associated with growth and reproduction traits of a large white pig and application of the CHRM3 gene InDel molecular marker, a 62bp deletion variation is identified in a second intron of the CHRM3 gene, and a breakpoint is positioned in a second intron 521426525142713bp (a pig chromosome 14 reference sequence NC010456.5) of ENSSSCT00000082112.1. The application for detecting / identifying the growth and reproduction traits of the large white pigs is developed on the basis of the molecular marker, and the detection method is simple, rapid and low in cost and can be used for accurately screening individuals with better traits such as backfat thickness, primary birth litter weight, total multiparous litter size and gestation period of multiparous sows in the early stage, so that genetic improvement is carried out on the growth and reproduction traits of the pigs, and the economic benefit is increased. Multiple characters such as growth and reproduction can be selected at the same time, the breeding period of excellent boars can be shortened, and the breeding efficiency and progress are accelerated.

The invention provides a single-chain and double-protein-expressed circular RNA construct and a preparation method and application thereof, and the single-chain and double-protein-expressed circular RNA construct comprises 5'and 3 'introns used for realizing RNA self-splicing and cyclization as a first kind of introns; the 5'and 3 'homologous arms are used as pairing sites in RNA molecules; the exon 1 and the exon 2, which are connected after being spliced, can be spliced and connected together under the action of the first type of introns to form a complete and continuous open reading frame; an interval sequence for providing spatial and adjustment sequence structures; the IRES is used for starting translation; a glycine-serine linker as a flexible linker; a green fluorescent protein for gene expression monitoring; a luciferase for producing light by a catalytic chemical reaction. All the advantages of the double-IRES system for generating natural complete protein are reserved, and the core defect of low efficiency can be thoroughly overcome.

Provided are a high-throughput transcriptome sequencing-based IKZF1 gene exon deletion recognition system and method. The exon deletion recognition system is obtained by inputting the number of IKZF1 exon junctions, differential genes and expression quantities thereof, and IKZF1 exon and intron mutation frequency information of samples as features into a constructed random forest model and performing prediction, and can be used for accurately predicting and recognizing any exon deletion with high accuracy and high specificity, providing guidance for patient prognosis and treatment.

The invention relates to an intron variant capable of accurately splicing and enhancing gene transcription and application of the intron variant, and belongs to the technical field of biological breeding, the intron variant is a variant of a Cat1mu intron, the Cat1mu intron has a nucleotide sequence as shown in SEQ ID NO.2, and the nucleotide sequence of the intron variant is formed by mutating base AG at the 182th to 183th sites of the sequence as shown in SEQ ID NO.2 into tG, Ac, At, cG, Aa, ta, tt, tc, cc, ct or ca. According to the present invention, the sequence analysis and the saturation mutation are performed on the Cat1mu, the screening is performed to obtain the Cat1mu series variant Cat1mu2-12, the accurate splicing effect of the Cat1mu2-12 during the fusion gene expression process is verified, and the Cat1mu6, the Cat1mu7, the Cat1mu8 and the Cat1mu10 can further enhance the fusion gene expression effect.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-I antigen gene, and belongs to the field of gene detection. The HLA-I antigen gene is subjected to PCR (Polymerase Chain Reaction) amplification by using 8 pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 7.8 Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, mixing samples, constructing an HLA-I type antigen gene library, carrying out accurate and complete sequencing on the full-length sequence of the HLA-I type antigen gene by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-I type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

The present invention discloses a circular gRNA and its related biomaterials and applications. The coding gene of the circular gRNA includes a 3' intron, a 3' splice site, a gRNA coding gene, a 5' splice site and a 5' intron in sequence, wherein the 3' splice site sequence is as shown in positions 2810-2814 of SEQ ID No.1, and the 5' splice site sequence is as shown in positions 2917-2930 of SEQ ID No.1. The circular gRNA is not attacked by nucleases, thereby improving the stability of the CRISPR / cas9 system.

The invention discloses a long-fragment PCR (Polymerase Chain Reaction) amplification reaction solution and application thereof. The PCR amplification reaction solution is composed of a 2XPCR buffer solution, dNTP, LA Taq enzyme, betaine and distilled water. In the PCR amplification reaction liquid, the volumes of the 2XPCR buffer solution, the dNTP, the LA Taq enzyme, the betaine and the distilled water in the total volume of 25l are respectively 12.5 l, 5.0 l, 0.25 l, 4.0 l and 0.45 l. According to the present invention, the PCR amplification reaction liquid can effectively amplify the long fragment PCR product, the product amplification with the longest detection reaching 12 kb can be used for the detection of the hemophilia A intron 22 inversion, the effective amplification reagent is provided for the long fragment PCR amplification, and the problem of difficult long fragment amplification is solved.

The application discloses a kind of molecular identification methods of sex-linked luhua feather homozygote of luhua chicken, belong to molecular marker assisted breeding technical field.The application can quickly and accurately detect homozygous sex-linked luhua chicken individual by designing KASP detection primer for detecting three sex-linked luhua feather color strong linkage sites, and the genotype of three sites can quickly and accurately detect homozygous sex-linked luhua chicken individual, when SNP2 (C->A) at 389bp of the first intron of CDKN2A gene, SNP3 and SNP4 at 172bp (T->A) and 174bp (T->C) of the first exon are AA, AA and CC respectively, then the chicken to be measured is homozygous sex-linked luhua chicken, otherwise the chicken to be measured is heterozygous sex-linked luhua chicken or non-sex-linked luhua chicken.The application provides an effective technical means for purifying sex-linked luhua feather color gene of luhua chicken, and helps to speed up the breeding speed of sex-linked luhua chicken strain homozygote and its matching system.

A method and compound for regulating SCN1A gene mRNA and protein expression. Provided is a use of a guide agRNA in preparation of a drug for altering expression of a target protein in a cell of a subject. The cell has a precursor mRNA encoding the target protein, wherein the precursor mRNA comprises an intron, an exon flanking a 5' splice site of the intron, and an exon flanking a 3' splice site or a pseudo 3' splice site of the intron; the guide agRNA is capable of binding to the precursor mRNA to form a structure capable of recruiting ADAR in the cell, thereby forming a complex containing an ADAR protein; and the target protein is a Nav1.1 protein. By regulating alternative splicing of a target gene pre-mRNA, the functional RNA of the target gene and the function or expression of the target protein are affected, thereby achieving the objective of disease treatment.

Abstract Methods for editing, e.g., introducing a heterologous transgene, within the human albumin gene (e.g., at intron 1) are provided. Abstract

The invention provides a method for plant transient expression of collagen and application of the plant transient expression of the collagen, and fragment collagen C3S1, C3S2, C3S3, C3S4 and C3S5 is formed by combining and connecting amino acid sequences shown as SEQ1, SEQ2, SEQ3, SEQ4 and SEQ5 respectively. According to the fragment collagen, plant tobacco can be used as a bioreactor, an expression vector is designed, the expression effect of the fragment collagen in the plant tobacco is compared, and meanwhile, a plant tobacco transient expression system is established, so that efficient expression of the fragment collagen in the tobacco is realized. By adopting the preparation method, tobacco can be utilized to express fragment collagen recombinant protein, a relatively high expression quantity is obtained, and compared with transformation by using full-length added intron or expression by using I-type collagen, the method realizes efficient expression of fragment collagen in III-type collagen in plant bioreactor tobacco, the period is short, and the yield is high. Maintenance of a cell line and a bioreactor is not needed, the production cost is remarkably reduced, and the safety index is high.

Lung cancer remains the leading cause of cancer-related deaths in the world. Despite the fact that introduction of immune checkpoint inhibitors (ICIs) led to a major advancement in lung cancer treatment, disease prognosis continues to remain low and a significant proportion of patients do not respond to such therapies. Cancer vaccines could potentially provide a complementary approach to boost antitumor immunity and act synergistically with ICIs. Novel tumor antigens shared by a large proportion of lung tumor cells are described herein. Several of the tumor antigens described herein derive from aberrantly expressed unmutated genomic sequences, such as intronic and intergenic sequences, which are not expressed in normal tissues. Nucleic acids, compositions, cells and vaccines derived from these tumor antigens are described. The use of the tumor antigens, nucleic acids, compositions, cells and vaccines for the treatment of lung cancer is also described.

The present invention relates to methods and devices for identification and quantification, including low abundance nucleotide base mutations, insertions, deletions, translocations, splice variants, miRNA variants, alternative transcripts, alternative start sites, alternative coding sequences, alternative non-coding sequences, alternative splicing, exon insertions, exon deletions, intron insertions, or other rearrangements and / or methylated nucleotide bases at the genomic level.

The application provides application of a SNP genetic marker affecting chicken body weight at the onset of lay in genetic breeding of laying hens, and belongs to the field of animal genetic breeding and biotechnology.The SNP genetic marker affecting chicken body weight at the onset of lay comprises bw1egg_1 and / or bw1egg_2; the Ensembl number of the bw1egg_1 is rs318020581, corresponding to the 76311052th position of the positive strand of chromosome 4 in the chicken reference genome bGalGal1.mat.broiler.GRCg7b sequence published in NCBI, belonging to the 3rd intron of the gene C1QTNF7, and the base at the position is T or G; the Ensembl number of the bw1egg_2 is rs313708699, corresponding to the 74731889th position of the positive strand of chromosome 4 in the chicken reference genome bGalGal1.mat.broiler.GRCg7b sequence published in NCBI, belonging to the 8th intron of the gene SLIT2, and the base at the position is T or C.Both the bw1egg_1 and the bw1egg_2 are helpful to genetically improve the body weight at the onset of lay, and when applied to genetic breeding of chickens, are favorable to improving the body weight at the onset of lay of the laying hens and obtaining a laying hen breed with excellent performance in uniformity of the body weight at the onset of lay.