500 results about "Intron" patented 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 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 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 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.

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.

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 invention discloses a general expression plasmid pHyg-HSRB-intron-3HA vector for Chlamydomonas reinhardtii as well as a construction method and application of the general expression plasmid pHyg-HSRB-intron-3HA vector. A nucleotide sequence of the general expression plasmid The vector comprises an HSRB promoter, an RBCS2intron, a target gene insertion site EcoRV, a 3 * HA tag at the 3'terminal, an rbcS2 terminator, a TUB2 promoter, a hygromycin B (Hyg +) screening gene and an rbcS2 terminator. The construction method comprises the following steps: constructing a skeleton of an expression plasmid pHyg-HSRB-intron-3HA vector; and constructing a combined fragment of an HSRB promoter and RBCS2intron, and carrying out homologous recombination on the combined fragment and a plasmid skeleton structure fragment to construct a vector for expressing plasmid pHyg-HSRB-intron-3HA. The method can improve the acquisition rate of positive transformants and the stable high expression of target genes.

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 streptomyces Targetron gene targeting vector as well as a construction method and application of the streptomyces Targetron gene targeting vector. According to the invention, an II-type intron Ll.LtrB from lactococcus lactis is cloned to an escherichia coli-streptomyces shuttle plasmid to construct a streptomyces genetic manipulation tool-vector pSC30 based on targeting of the II-type intron, and gene targeting in streptomyces coelicolor is successfully realized by using the tool. Screening a biosynthetic gene cluster of red pigment in streptomyces roseosporus; and activating a biosynthetic gene cluster of janus doxorubicin in streptomyces spinosus. According to the invention, the conversion rate and the gene targeting efficiency of the Ll.LtrB intron in streptomyces with high GC content are effectively improved, and a new tool is provided for construction of a gene mutation library.

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 relates to the technical field of gene vectors, and discloses a multi-target multi-gene stable silencing vector and an application thereof in treating diabetes mellitus, and a multi-gene multi-target gene modification combined element is safely and efficiently integrated at a gene safety parking point of a second intron region of a CCR5 gene in a genome in a reverse fixed point manner by using a non-viral vector system; therefore, the effect of continuously silencing expression of a plurality of target genes such as MSTN, GRB10 and PTBP1 for a long time is achieved, and aging of mesenchymal stem cells cultured in vitro is remarkably inhibited. When the mesenchymal stem cells modified by the multi-target multi-gene stable silencing vector are infused, the activity is high, the chronic inflammatory microenvironment is effectively inhibited, and the senescence process is delayed; moreover, proliferation and differentiation of pancreatic beta cells are promoted, insulin secretion is enhanced, insulin resistance is effectively resisted, and diseases such as diabetes mellitus are prevented.

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 discloses a universal and'scar '-free RNA cyclization method, which comprises a recombinant nucleic acid molecule for preparing circular RNA, the recombinant nucleic acid molecule comprises the following elements arranged in sequence along the direction from 5'to 3': an optional 5 'homologous arm, a 3' semi-intron fragment, a cyclization fragment, a 5 'semi-intron fragment and an optional 3' homologous arm; the 5'end of the cyclization fragment comprises a 3 'cyclization recognition fragment, and the 3' end of the cyclization fragment comprises a 5 'cyclization recognition fragment.