95 results about "DNA construct" patented technology

The present disclosure relates to the seminal discovery of a generation and use of genetically engineered Vibrio sp. Provided is the use of the genetically engineered bacteria for the construction, maintenance, manipulation, and / or propagation of DNA constructs; protein expression; protein secretion; vectors and other metabolic tools; metabolic engineering; expression of cellular extracts for cell-free biology; shuttle vectors; cloning vectors; and for synthetic biology applications. The disclosure also relates to the use of the replication machinery of Vibrio sp. as a cloning or expression vector for replication of recombinant DNA constructs. The disclosure also relates to methods of use of the above.

The invention relates to a method to determine a homology directed repair (HDR) event within a eukaryotic cell, wherein the cell expresses a first isoform of a surface protein, which is different from a second isoform of said surface protein with regard to an amino acid marker. The method comprises the steps of inducing a DNA double strand break, providing a HDR template DNA construct comprising the amino acid marker corresponding to the second isoform of the surface protein and subsequently determining the expression of the first or second isoform of said surface protein on said cell, wherein expression of the second isoform indicates a successful HDR event. The invention also relates to a method for editing a genomic location of interest within a eukaryotic cell, and to a method of selectively depleting or enriching an edited cell in a composition of non-edited and edited cells.

Compositions and methods for conferring pesticidal activity to bacteria, plants, plant cells, tissues and seeds are provided. Compositions comprising a coding sequence for a toxin polypeptide are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also comprise transformed bacteria, plants, plant cells, tissues, and seeds. In particular, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are encompassed, and antibodies specifically binding to those amino acid sequences. In particular, the present invention provides for isolated nucleic acid molecules comprising nucleotide sequences encoding the amino acid sequence shown in any of SEQ ID NO: 16 to 30, or the nucleotide sequence set forth in any of SEQ ID NO: 1 to 15, as well as variants and fragments thereof.

Provided herein are methods and compositions for truncating and, optionally, joining polynucleotides. The methods bypass inefficient cloning methods and the requirement for expensive enzymes by using in vivo homologous recombination in conjunction with specific endonuclease digestion. The methods may further be used to assemble long fragments of DNA from existing DNA constructs or libraries or to generate novel DNA libraries.

The present invention provides a simple and inexpensive system for regulating signal peptide performance by using a synthetically evolved nucleotide sequence. The invention further relates to an expression vector comprising the nucleotide sequence. Additionally, the present invention relates to host cell comprising the expression vector. Furthermore, the present invention relates to a recombinant protein expressed by the host cell as well as a method for expressing the recombinant protein.

The present invention relates to a DNA construct that codes for a fusion protein comprising at least two enterohemorrhagic Escherichia coli antigens, for example, the antigens Int280, EspB, EspA, Stx2B, EspD, combinations or fragments thereof, vectors, fusion protein, immunisation methods and vaccines. The coded antigens can be fused Int280 and EspB of enterohemorrhagic Escherichia coli, forming a fusion protein with or without a connecting sequence between the two. The fused antigens can show the amino acid sequence SEQ ID No. 3 or SEQ ID No. 4 and are coded by the nucleotide sequence in SEQ ID No. 1 and SEQ ID No. 2. The construct can also comprise a signal sequence and a binding sequence for binding to the membrane of bacteria.

Cyclic dinucleotides (CDRs) are signaling molecules for cancer therapy, acting as second messengers for intracellular events initiated by GPCR activation and stimulating factors for the interferon gene (STING), thus significantly influencing tumor suppression. Simultaneously, when cancer occurs in a subject, angiogenesis and cell growth proceed at very high rates in vivo, creating a hypoxic environment due to incomplete angiogenesis in cancerous tissue. This environment can be highly conducive to the proliferation of anaerobic bacteria such as Salmonella strains or Escherichia coli. Therefore, this invention relates to a DNA construct incorporating a gene encoding an enzyme for synthesizing cancer therapeutic signaling molecules (e.g., CDRs), or to strains transformed with a vector containing the DNA construct. According to the invention, the DNA construct or strains transformed with a vector containing the DNA construct target cancer in a subject and then secrete C-di-AMP or C-di-GMP synthases in the surrounding environment of the cancer, thus enabling highly effective prevention or treatment of cancer while allowing for real-time cancer diagnosis.

In one aspect, disclosed herein are DNA constructs and biological devices comprising S. cerevisiae cells and extracts and lysates produced therefrom, wherein the DNA constructs encode genes related to NAD+ synthesis and wherein the extracts and lysates produced therefrom include one or more of NAD+, NADP+, NADH, and / or NADPH. Also disclosed are nutritional and pharmaceutical compositions including the extracts and lysates and the coenzymes produced therefrom. In some aspects, the disclosed extracts and lysates can be added to food and beverage products to improve the nutritional profiles of the same.

Compositions and methods for imparting insecticidal activity to bacteria, plants, plant cells, tissues, and seeds are provided. Compositions comprising coding sequences for toxin polypeptides are provided. The coding sequences can be used in DNA constructs or expression cassettes for transformation and expression in plants and bacteria. Compositions also include transformed bacteria, plants, plant cells, tissues, and seeds. Specifically, isolated toxin nucleic acid molecules are provided. Additionally, amino acid sequences corresponding to the polynucleotides are contemplated, and antibodies specifically bind to those amino acid sequences. Specifically, the present invention provides isolated nucleic acid molecules comprising a nucleotide sequence encoding the amino acid sequence set forth in any one of SEQ ID NOs: 16 to 30 or the nucleotide sequence set forth in any one of SEQ ID NOs: 1 to 15, as well as variants and fragments thereof.

The present invention is generally in the field of synthetic biology, gene therapy and cell therapy. In particular, the present invention relates to synthetic transcription factors comprising a DNA binding domain derived from a mitochondrial DNA binding protein (e.g., MTERF1) and a transcriptional regulatory domain derived from one or more other proteins; nucleic acids or combinations of nucleic acids encoding the synthetic transcription factors of the invention; a DNA construct comprising an MTERF1 binding site and a minimum promoter; a system comprising a synthetic transcription factor of the invention and a DNA construct of the invention; and uses thereof, such as medical uses.

Compositions and methods for modifying genomic DNA sequences are provided. The methods produce double-stranded breaks (DSBs) at pre-determined target sites in a genomic DNA sequence, resulting in mutation, insertion, and / or deletion of DNA sequences at the target site(s) in a genome. Compositions comprise DNA constructs comprising nucleotide sequences that encode a Cms1 protein operably linked to a promoter that is operable in the cells of interest. The DNA constructs can be used to direct the modification of genomic DNA at pre-determined genomic loci. Methods to use these DNA constructs to modify genomic DNA sequences are described herein. Additionally, compositions and methods for modulating the expression of genes are provided. Compositions comprise DNA constructs comprising a promoter that is operable in the cells of interest operably linked to nucleotide sequences that encode a mutated Cms1 protein with an abolished ability to produce DSBs, optionally linked to a domain that regulates transcriptional activity. The methods can be used to up- or down-regulate the expression of genes at predetermined genomic loci.

The present disclosure provides recombinant polypeptides containing a payload sequence and one or more 3'-untranslated region (3'-UTR) sequences downstream of and operably linked to the payload sequence. The one or more 3'-UTR sequences, or RNA transcripts thereof, respond to an intracellular signal by modulating the translation, transcription, and / or stability of the payload sequence or an RNA transcript thereof. The disclosure also provides DNA constructs, vectors, cells, and systems including the provided recombinant polypeptides, as well as methods for modifying cells and / or preventing or treating a disease by using the provided materials.

Described herein are genelight cultures, extracts and related methods. In one aspect, the method of making a genelight culture or extract includes the steps of (a) making a DNA construct containing genes for producing a heat shock protein, RuBisCO large subunit 1, tonB, hydrogenase, and a P-type ATPase, (b) introducing the DNA construct into host microbial cells via transformation or transfection, and (c) culturing the microbial cells to produce the genelight cultures and extracts. The compositions of these cultures and extracts can be tailored to have specific properties such as the ability to provide power to a light emitting diode. The cultures and extracts have further uses including enhancing the growth of plants and as supplemental nutrients of cultures of industrially important microorganisms. The cultures and extracts further have UV-protective properties. Also described herein are microbial electric circuits containing the cultures and extracts described herein.

The present disclosure relates to DNA constructs encoding a CD19-specific chimeric antigen receptor (CAR) and other recombinant proteins, for example, a CD70-binding protein or a CD70-specific CAR, and engineered immune cells (e.g., CD19-specific CAR T cells or CD19 / CD70 dual CAR T cells) comprising the DNA constructs, methods of making the same, and uses thereof in therapeutic applications.

A pesticidal protein class exhibiting toxic activity against Coleopteran and Lepidopteran pest species is disclosed, and includes, but is not limited to, TIC7040, TIC7042, TIC7381, TIC7382, TIC7383, TIC7386, TIC7388, and TIC7389. DNA constructs are provided which contain a recombinant nucleic acid sequence encoding the TIC7040, TIC7042, TIC7381, TIC7382, TIC7383, TIC7386, TIC7388, and TIC7389 pesticidal proteins. Transgenic plants, plant cells, seed, and plant parts resistant to Coleopteran and Lepidopteran infestation are provided which contain recombinant nucleic acid sequences encoding the TIC7040, TIC7042, TIC7381, TIC7382, TIC7383, TIC7386, TIC7388, and TIC7389 pesticidal proteins of the present invention. Methods for detecting the presence of the recombinant nucleic acid sequences or the proteins of the present invention in a biological sample, and methods of controlling Coleopteran and Lepidopteran species pests using the TIC7040, TIC7042, TIC7381, TIC7382, TIC7383, TIC7386, TIC7388, and TIC7389 pesticidal proteins are also provided.

Pesticidal proteins exhibiting inhibitory, suppressive, and toxic activity against Lepidopteran pest species are disclosed, and include, but are not limited to, TIC4064 and TIC4064 amino acid sequence variants. DNA constructs are provided which contain a recombinant nucleic acid sequence encoding one or more of the disclosed pesticidal proteins. Transgenic plants, plant cells, seed, and plant parts resistant to Lepidopteran infestation are provided which contain recombinant nucleic acid sequences encoding the pesticidal proteins of the present invention. Methods for detecting the presence of the recombinant nucleic acid sequences or the proteins of the present invention in a biological sample, and methods of controlling Lepidopteran species pests using any of the TIC4064 and TIC4064 amino acid sequence variant pesticidal proteins are also provided.

A novel ubiquitous chromatin opening element (UCOE) named SRF-UCOE and methods for its use are provided. Compositions including recombinant and synthetic SRF-UCOE nucleic acid molecules, DNA constructs and vectors comprising the SRF-UCOE nucleic acid molecules, host cells comprising the DNA constructs or vectors, and cell culture systems comprising such host cells. SRF-UCOE polynucleotide sequences can be used in DNA constructs or expression cassettes for transformation and expression in cells or organisms of interest. The compositions and methods provided are useful for increasing and / or maintaining expression of a gene of interest. Transgenic cells, tissues, and animals comprising a SRF-UCOE nucleotide sequence are also provided. Methods are provided for increasing and / or maintaining expression of a gene of interest and for treating a subject via gene therapy.

The present invention relates to means and methods for producing adeno-associated virus capsid libraries using insect host cells. In particular, the present invention relates to novel DNA constructs and methods of using these constructs to produce libraries of adeno-associated virus capsids in insect host cells, which minimize the occurrence of cross-packaging and chimeric phenomena in these libraries produced. The present invention further relates to methods for identifying AAV capsid variants having one or more desired characteristics such as, for example, a combination of CNS tropism and peripheral organ de-targeting.

Compositions having pesticidal activity and methods for their use are provided. Compositions include isolated and recombinant polypeptide sequences having pesticidal activity, recombinant and synthetic nucleic acid molecules encoding the pesticidal polypeptides, DNA constructs comprising the nucleic acid molecules, vectors comprising the nucleic acid molecules, host cells comprising the vectors, and antibodies to the pesticidal polypeptides. Nucleotide sequences encoding the polypeptides provided herein can be used in DNA constructs or expression cassettes for transformation and expression in organisms of interest. The compositions and methods provided herein are useful for the production of organisms with enhanced pest resistance or tolerance. Transgenic plants and seeds comprising a nucleotide sequence that encodes a pesticidal protein of the invention are also provided. Methods are provided for producing the polypeptides disclosed herein, and for using those polypeptides for controlling a pest. Methods and kits for detecting polypeptides of the invention in a sample are also included.

The present invention relates to a method for editing the genome of a cell, such as a yeast cell. The method of the present invention requires the cell to be contacted with at least one ribonucleoprotein, at least one donor-DNA construct and a selectable marker such that they are introduced into the cell. The present invention is especially suitable for multiplex genome editing of cells such as yeast cells. The current invention further relates to a composition, a cell obtainable by the method of the invention and a method for the production of a compound of interest.

A DNA construct for expressing a recombinant protein, wherein the DNA construct comprises: —at least one of the nucleotide sequences of SEQ ID No 20 and 21, wherein a nucleotide sequence of SEQ ID 20 and 21 is a TIR sequence; and —a nucleotide sequence which encodes a signal peptide; and wherein a nucleotide sequence of SEQ ID No 20 and 21 comprises at least the first 9 nucleotides of said signal peptide encoding sequence.

The present disclosure provides methods and compositions for improving the in vitro transcription (IVT) of messenger RNA (mRNA) using SP6, T7, or KP34 RNA polymerase. In particular, the present disclosure provides in vitro transcription templates with optimized nucleic acid sequences specific for use with SP6, T7, or KP34 RNA polymerase that improve mRNA transcript yield and / or reduce amounts of dsRNA.

This disclosure relates to devices and methods for administering therapeutic compositions to the pharyngeal muscles. Specifically, the devices and methods of this disclosure may be used to treat or prevent dysphagia, including dysphagia associated with oculopharyngeal dystrophy (OPMD), in individuals suffering from or predisposed to dysphagia. In addition, this disclosure relates to devices and methods used to deliver modified adeno-associated virus (AAV) delivery vectors containing a "silence and substitution" DNA construct, including compositions containing a "silence and substitution" DNA construct.

Provided herein are variant Cas12a proteins comprising at least one artificial evoked mutation. Also provided are fusion proteins comprising these variant Cas12a proteins and one or more heterologous domains. Related nucleic acids, DNA constructs, vectors, cells, and methods of editing nucleic acids using these variant Cas12a proteins and / or fusion proteins are also provided. Using the provided proteins can increase the frequency of desired nucleic acid editing (e.g., SDN-1 editing in the plant genome).

The present invention relates to a DNA construct comprising: a gene encoding flagellin and a gene encoding an immunopotentiator (or adjuvant) protein. In order to achieve effective cancer treatment by selectively killing only cancer cells, the attenuated Salmonella strains of the present disclosure are designed to produce immunogenic substances in cancer tissue to induce a strong anti-cancer immune response to significantly inhibit tumor size in primary and metastatic cancers. Therefore, the strain can be advantageously used in a prophylactic or therapeutic composition for increasing survival rate.

This application relates to a DNA construct for overexpressing a target gene in *Schizochytrium* microorganisms, and a method for expressing the target gene in *Schizochytrium* microorganisms using the construct. The DNA construct includes a 5' homologous arm and a 3' homologous arm capable of hybridizing to a high-expression site in the *Schizochytrium* microorganism genome.

The present disclosure provides nucleic acids obtained from Bacillus thuringiensis strains, and variants and fragments thereof, which encode polypeptides having pesticidal activity against insect pests including Lepidoptera. Particular embodiments of the present disclosure provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms and plants comprising the nucleic acids of the embodiments. These compositions can be applied in methods for controlling pests, particularly plant pests.

The invention provides nucleic acids, and variants and fragments thereof, obtained from strains of Bacillus thuringiensis encoding polypeptides having pesticidal activity against insect pests, including Lepidoptera. Particular embodiments of the invention provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms. These compositions find use in methods for controlling pests, especially plant pests..

The present invention provides loop-mediated riboregulators for the detection of SARS-CoV-2. Also provided are DNA constructs encoding the loop-mediated riboregulators and methods of using the loop-mediated riboregulators to detect the presence of SARS-CoV-2 in a sample.