42 results about "Poly-ADP-Ribosylation" patented technology

One-step RT-PCR methods, compositions and kits for the detection and quantification of small RNAs in a sample are disclosed. The one-step RT-PCR approach involves polyadenylation of a small RNA followed by reverse transcription with a first primer containing a poly(T) sequence and at least two 3′ nucleotides complementary to the 3′ terminal end nucleotides of the small RNA, to produce a cDNA. This may be followed by PCR amplification using the same first primer as the revere primer and a second, forward primer in which a portion of its sequence is complementary to the 3′ terminal end of the cDNA. This may be then followed by detection and/or quantification of the amplified product.

Methods and means are provided for reducing the phenotypic expression of a nucleic acid of interest in eukaryotic cells, particularly in plant cells, by providing aberrant, preferably unpolyadenylated, target-specific RNA to the nucleus of the host cell. Preferably, the unpolyadenylated target-specifc RNA is provided by transcription of a chimeric gene comprising a promoter, a DNA region encoding the target-specific RNA, a self-splicing ribozyme and a DNA region involved in 3′ end formation and polyadenylation.

The human soluble neuropilin-1 (sNRP) polyadenylation signal (sNRP-poly(A)), situated downstream of the GT splice donor site of intron 12 of the full-length neuropilin-1 gene, also functions as the termination codon for sNRP. This 17 nucleotide sequence efficiently facilitates addition of poly(A) tails to RNAs expressed in cells. The present invention shows that this optimally succinct sequence has similar activity to the SV40 polyadenylation signal that is currently used in expression vectors. By using this shorter dual termination/polyadenylation signal and avoiding the need for large and cumbersome polyadenylation signals, expression vectors may be engineered to carry considerably larger genes.

The present invention provides novel isolated nucleic acids that comprise an avian nucleic acid sequence comprising a ovomucoid gene expression control region. The ovomucoid promoter region of the present invention will allow expression of an operably linked heterologous nucleic acid insert in a transfected avian cell such as, for example, an oviduct cell. The isolated avian ovomucoid of the present invention may be operably linked with a selected nucleic acid insert, wherein the nucleic acid insert encodes a polypeptide desired to be expressed in a transfected cell. The recombinant DNA of the present invention may further comprise a polyadenylation signal sequence. The present invention further includes expression vectors comprising an isolated avian ovomucoid gene expression control region of the present invention, and transfected cells and transgenic avians comprising the expression vectors.

Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5′ promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5′-terminus of the coding region for the second protein to form a construct for expression of more than two proteins. A single 3′ termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and/or value added products, illustrated by the production of polyhydroxyalkanoates in plants.

A method for preparing an electrochemical sensor for non-coding RNA, which comprises preparing a sulfonated cup [8] arene-supported reduced graphene oxide by a one-step method, synthesizing a ferroferric oxide nano material by a moist heating method, and adding gold nanoparticles to synthesize Au@SCX8-RGO and Au@Fe3O4 composite materials. After confirming successful material construction by material characterization, a target material is electrochemically detected by a screen printing electrode with toluidine blue and other materials used as electrical signal substances. The electrochemical biosensor prepared by the method can detect the expression quantity of 3'UTR transcripts of non-coding regions of different lengths produced by the selective polyadenylation (APA) phenomenon.

The present invention relates to a genic expression adenoviral hybrid vector characterized in that it contains at least the following elements, oriented in the direction 5′ to 3′: i. a first chain of adenoviral origin comprising a first inverted terminal repeat (ITR) sequence and a signal sequence for packaging of the adenovirus; ii. a first non-encoding stuffer sequence; iii. a sequence corresponding to a tissue specific promoter; iv. a chain of cDNA derived from an alphavirus, the sequence of which is partly complementary to an alphaviral RNA sequence, comprising at least a sequence encoding for at least one exogenous gene of interest; v. a polyadenylation sequence; and vi. a second adenoviral inverted terminal repeat (ITR) sequence, it preferably relates to an adenoviral hybrid vector comprising as exogenous gene of interest the therapeutic gene of mammalian interleukin IL-12 and even more preferably human interleukin hIL-12; and to the use of the hybrid vector in a process for transferring genetic material to a cell, particularly a tumor cell that preferably expresses alpha-fetoprotein (AFP), and to its use for inducing an immune response against foreign antigens.

Disclosed are materials and methods for creating customizable traits in animals. In the demonstration of the principle of the subject invention, a keratin-14 specific promoter is used with red fluorescent protein in the loxp cassette, dominant black ([Delta]G23) beta defensin 103 in the pigment cassette, and an SV40 (with intron) polyadenylation sequence. When Cre recombinase (or HTNCre) is applied to the animal's skin in a carrier base (e.g., lipid bilayers), fur is permanently genetically modified to tum black in the shape in which theHTNCre was applied.

The present invention provides methods for designing a sequence for efficient short interference RNA molecules. In particular, the present invention defines a universal target for siRNA derived from the consensus sequence of the polyadenylation signal in conjunction with unique sequences for gene silencing and inhibition of viral replication in a eukaryotic host cell. The present invention further provides methods for the treatment and prevention of diseases and disorders by silencing a gene of a virus, an oncogene, genes encoding transcription factors and many other diseases related genes.

The invention relates to application of cordycepin in in-vitro maturation culture of oocytes of small pig follicles, which belongs to the field of auxology of animals. Degrees of polyadenylation of the maternal genes consisting of cyclinB1, C-mos, GDF9, BMP15 and Cdc2 in oocytes determine expression levels of the genes, which has certain promotion effects on the recruitment process where small follicles become dominant follicles and on maturation of oocytes. The expression levels of the genes consisting of C-mos, GDF9 and BMP15 play a leading role in maturation of the oocytes of small follicles. Addition of a 3'-dA inhibitor can improve the expression levels of the genes consisting of C-mos, GDF9 and BMP15, so the maturation rate of the small follicles is increased. The technology of in-vitro maturation culture of oocytes is expected to be popularized and applied in research fields like transgenic technology, nuclear transplantation technology and embryonic stem cell technology.

An isolated DNA molecule having a sequence which comprises in a 5′ to 3′ direction (i) one or more promoter elements, (ii) the geneof interest, and (iii) a poly-adenylation 5 signal, and (iv) a terminator element, and expressing the geneof interest incorporated into the DNA molecule in an expression system, and use of said molecule to enhance expression of a gene of interest.

The invention relates to compositions and methods to isolate nucleic acids, and the identification of polyadenylation sites in a gene of interest. In one aspect, the invention provides an oligonucleotide comprising at least one nucleic acid and an affinity moiety, wherein said nucleic acid is 30-60 nucleotides in length and said nucleic acid comprises 1-25 uracil and 5-50 thymine nucleotides.

The invention discloses an expression vector expressing a recombinant complete HBV surface antigen comprising L protein, M protein and S protein, prepared by injecting a polynucleotide of a coding region which has a complete HBV envelope gene and a complete 3'-UTR nucleotide containing polyadenylation sites into a pSGM vector identified by the preservation number KCCM 10202.

The present invention provides genomic and cDNA encoding human cytoplasmic polyadenylation element binding protein, expression vectors comprising human cytoplasmic polyadenylation element binding protein cDNA and host cells that contain the expression vectors. Also provided are recombinant human cytoplasmic polyadenylation element binding protein and polypeptides derived thereof. In addition, the present invention reports that the human Mos 3′ untranslated region (3′ UTR) contains a functional cytoplasmic polyadenylation element (CPE), interacts with the human CPEB1 protein and directs maturation-dependent cytoplasmic polyadenylation of the endogenous Mos mRNA.

A recombinant adenovirus expressing a streptolysin O (SLO) protein comprising a SLO gene; a promoter operably linked to the SLO gene; a polyadenylation signal sequence; and an adenovirus genome lacking E1 gene effectively kills a cancer cell by expressing a pore-forming toxin, SLO protein, and, therefore, is useful for the suicide cancer gene therapy.

Disclosed are a pharmaceutical composition for treatment of neurodegenerative diseases or diseases caused by abnormality of RNA binding protein and applications thereof, in particular the application in the treatment of ALS. The pharmaceutical composition can significantly enhance the dynamic performance of stress particles containing RNA binding proteins such as hnRNP A1 and TDP-43 proteins; influences the interaction between the RNA binding proteins and other poly ADP ribosylation modified proteins or other PAR binding proteins; influences the subcellular localization and stress response of RNA binding proteins; influences the liquid-liquid phase separation and aggregation tendency of RNA binding proteins; influences the co-phase separation between RNA binding proteins; influences the interaction of RNA binding proteins in cells; and has a significant inhibitory effect on neurotoxicity caused by RNA binding proteins.

The invention discloses a cytoplasmic polyadenylation element-binding protein 4 vaccine, and preparation and application thereof. The vaccine is a ISCOM-DNA compound, the preparation method of the vaccine comprises: cloning of full-length DNA of cytoplasmic polyadenylation element-binding protein 4, cloning of full-length DNA of tumor necrosis factor BAFF, construction of a recombinant vector pStar-CPEB4/BAFF, construction of a pStar-CPEB4/BAFF engineering bacteria, inducible expression of CPEB4, and preparation of the ISCOM-DAN compound. The vaccine has good application prospect on neuroglioma controlling. The vaccine has the advantages of being efficient, low in toxicity, simple in preparation method and the like, and is applicable to medicines for tumour immunotherapy.

The present invention provides a linear expression construct free of any conventional amplification and/or selection sequences comprising an RNA polymerase I (polI) promoter and a polI termination signal, inserted between a RNA polymerase II (polII) promoter and a polyadenylation signal useful for the expression of segments of viral RNA, preferably influenza viruses. The inventive construct is useful for efficient and fast production of viral particles, especially for producing vaccine formulations for the treatment of epidemic and/or pandemic diseases.

The present invention provides an integration-defective lentiviral vector based on a parental lentivirus and related methods, the integration-defective lentiviral vector including one or more of the following: (a) a mutation, deletion or other modification of one or more binding sites for a host factor involved in gene silencing; (b) an addition of one or more binding sites for a transcription activator, which can be natural (such as but not limited to ubiquitous and/or tissue/cell type specific) including but not limited to SP1 NFkB, or synthetic including but not limited to binding sites for tetracycline regulated trans activators tTA, rtTA, tT65, and/or rtT65; (c) one or more nucleic acid sequences from a SV40 genome, wherein the one or more sequences are obtained from a region of the SV40 genome upstream to the SV40 poly-adenylation signal; (d) a shRNA expression cassette, which encodes a shRNA directed to a host gene involved in epigenetic silencing and/or in DNA repair pathways; or (e) any combination of (a), (b), (c) and (d), wherein as compared to the parental lentivirus, the integration-defective lentiviral vector resists gene silencing.