Abstract: This disclosure relates to modified single and dual guide RNAs having improved in vitro and in vivo activity in gene editing methods.

Abstract: The present invention describes compositions and methods for priming protective immunity in the presence of pre-existing maternal antibody. In some embodiments, the invention contemplates simultaneously masking vaccines to avoid antibody neutralization while targeting those vaccines to specific cell types in order to elicit an enhanced immune response. In other embodiments, vectors that recruit and activate specific antigen-presenting cells may further enhance the efficacy of those immune responses.

Abstract: The present disclosure provides conditionally active, heterodimeric polypeptides. The conditionally active, heterodimeric polypeptides are active in the presence of a dimerizing agent that induces dimerization of the polypeptides of the heterodimer. A conditionally active, heterodimeric polypeptide of the present disclosure is useful in a variety of research and treatment methods, which are also provided.

Abstract: The disclosure is directed to methods, kits, and compositions for amplifying and detecting a human immunodeficiency virus-1 (HIV-1) in a sample, which comprises a variety of combinations of forward oligonucleotide primers, reverse oligonucleotide primers, and oligonucleotide probes.

Abstract: Methods and compositions for bacterial production of pure single-stranded DNA (ssDNA) composed of custom sequence and size have been developed. The methods enable scalability and bio-orthogonality in applications of scaffolded DNA origami, offering one-step purification of large quantities of pure ssDNA amendable for immediate folding of DNA nanoparticles. The methods produce pure ssDNA directly from bacteria. In some embodiments the E. coli helper strain M13cp combined with a phagemid carrying only an f1-origin allows for, without the need for additional purification from contaminating dsDNA. This system is useful for generalized circular ssDNA synthesis, and here is applied to the assembly of DNA nanoparticles folded both in vitro and direct from phage.

Abstract: The present invention describes compositions and methods for priming protective immunity in the presence of pre-existing maternal antibody. In some embodiments, the invention contemplates simultaneously masking vaccines to avoid antibody neutralization while targeting those vaccines to specific cell types in order to elicit an enhanced immune response. In other embodiments, vectors that recruit and activate specific antigen-presenting cells may further enhance the efficacy of those immune responses.

Abstract: The invention relates to retrieving or modifying target nucleic acids, such as host cell chromosomal DNA, by homologous recombination with vectors that contain homology arms flanked by nuclease cutting sites that have been cut by a nuclease such as Cas to provide recombinogenic nucleic acid strands in situ. The methods described herein can be used in genetic engineering of cells and for gene therapy of cells or organisms.

Abstract: The disclosure generally relates to compositions and methods for the production of nucleic acid molecules. In some aspects, the invention allows for the microscale generation of nucleic acid molecules, optionally followed by assembly of these nucleic acid molecules into larger molecules. In some aspects, the invention allows for efficient production of nucleic acid molecules (e.g., large nucleic acid molecules such as genomes).

Abstract: Disclosed are compositions including primers and probes, which are capable of interacting with the disclosed nucleic acids, such as the nucleic acids encoding the reverse transcriptase or protease of HIV as disclosed herein. Thus, provided is an oligonucleotide comprising any one of the nucleotide sequences set for in SEQ ID NOS:1-89, and 96-104. Also provided are the oligonucleotides consisting of the nucleotides as set forth in SEQ ID NOS:1-89, and 96-104. Each of the disclosed oligonucleotides is a probe or a primer. Also provided are mixtures of primers and probes and for use in RT-PCR and primary PCR reactions disclosed herein. Provided are methods for the specific detection of several mutations in HIV. Mutations in both the reverse transcriptase and the protease of HIV can be detected using the methods described herein.

Abstract: The present invention describes compositions and methods for priming protective immunity in the presence of pre-existing maternal antibody. In some embodiments, the invention contemplates simultaneously masking vaccines to avoid antibody neutralization while targeting those vaccines to specific cell types in order to elicit an enhanced immune response. In other embodiments, vectors that recruit and activate specific antigen-presenting cells may further enhance the efficacy of those immune responses.

Abstract: The invention provides adeno-associated virus (AAV) replication (Rep) sequences. In one embodiment, the invention provides nucleotide sequences encoding a chimeric protein, wherein the encoded chimeric protein contains a wild type AAV Rep inhibitory amino acid sequence, and wherein the nucleotide sequences contain a scrambled and/or deoptimized polynucleotide sequence encoding the wild type AAV Rep inhibitory amino acid sequence. The invention provides vectors, cells, and viruses containing the invention's sequences. Also provided are methods for detecting portions of the AAV Rep inhibitory amino acid sequence, which reduce replication and/or infection and/or productive infection by viruses. The invention's compositions and methods are useful for site-specific integration and/or expression of heterologous sequences by recombinant adeno-associated virus (rAAV) vectors and by rAAV virus particles, such as hybrid viruses (e.g., Ad-AAV) comprising such vectors.

Abstract: A cell culture system for fermentation or cultivation of at least one of cells, tissues or tissue-like cell cultures, organs or organ-like cell cultures and multicellular organisms comprises: a system vessel (1) in which at least one cavity (6) is formed, the cavity (6) having at least three ports (2, 3, 4); at least one porous substrate (9) installed within the cavity (6) so as to allow fluid flowing from any one to any other of said three ports (2, 3, 4) to pass through said porous substrate (9); and, at least one channel extending from a first one of said ports (3) within the porous substrate (9) towards a central region of said porous substrate (9).

Abstract: The present invention relates to a vector system involving replacement of a Woodchuck Hepatitis Virus Post-Transcriptional Regulatory Element (WPRE) sequence with an unrelated short spacer sequence for efficient expression of nucleotides of interest in a retroviral vector system and methods of delivering and expressing nucleotides of interest to target cells.

Abstract: An isolated fungus is described. The isolated fungus produces at least one compound selected from the group consisting of 1,8-cineole, 1-methyl-1, 4-cyclohexadiene, and (+)-?-methylene-?-fenchocamphorone. A method for producing at least one compound selected from the group consisting of 1,8-cineole, 1-methyl-1, 4-cyclohexadiene, and (+)-?-methylene-?-fenchocamphorone is also described. The method includes culturing a fungus on or within a culturing media in a container under conditions sufficient for producing the at least one compound.

Abstract: Anti-angiogenic adenovirus vectors, and therapeutic use thereof are provided, and more particularly, but not exclusively, clinical protocols for treatment of solid tumors in patients with an Ad5-PPE-1-3X-fas-chimera adenovirus vector.

Abstract: Complex viruses are assembled from simple protein subunits by sequential and irreversible assembly. During genome packaging in bacteriophages, a powerful molecular motor assembles at the special portal vertex of an empty prohead to initiate packaging. An aspect of the invention relates to the phage T4 packaging machine being highly promiscuous, translocating DNA into finished phage heads as well as into proheads. Single motors can force exogenous DNA into phage heads at the same rate as into proheads and phage heads undergo repeated initiations, packaging multiple DNA molecules into the same head. This shows that the phage DNA packaging machine has unusual conformational plasticity, powering DNA into an apparently passive capsid receptacle, including the highly stable virus shell, until it is full. These features allow for the design of a novel class of nanocapsid delivery vehicles.

Abstract: A method of determining sensitivity or resistance of isolates of HIV retroviruses to a molecule includes a) amplifying sequences coding for a protease of a retrovirus to be studied, with or without the or some of amino acid sequences situated upstream and downstream of a cleavage site of a precursor in which the amino acid sequences are situated, b) recombining fragments of DNA, a final product of the amplification, and an expression vector allowing expression of sequence coding for the protease of the retrovirus to be studied under control of a known inducible promoter through co-transformation of the vector and the DNA fragments with at least one yeast cell, c) culturing co-transformed yeast cell or cells to obtain a sufficient number of transformants to perform a sensitivity or resistance test, and recovering transformants issuing from the co-transformed cell, on any suitable medium, d) incubating the transformants in the presence of a molecule to be tested, e) qualitatively or quantitatively analyzing the

Abstract: An in vitro process for the production of closed linear deoxyribonucleic acid (DNA) comprises (a) contacting a DNA template comprising at least one protelomerase target sequence with at least one DNA polymerase in the presence of one or more primers under conditions promoting amplification of said the template; and (b) contacting amplified DNA produced in (a) with at least one protelomerase under conditions promoting production of closed linear DNA. A kit provides components necessary in the process.

Abstract: A method for producing at least one compound selected from the group consisting of 1,8-cineole, and 1-methyl-1,4-cyclohexadiene is also described. The method includes culturing a microorganism on or within a culturing media in a container under conditions sufficient for producing the at least one compound.

Abstract: An expression vector capable of expressing a foreign gene in Pseudonocardia autotrophica; a transformant of Pseudonocardia autotrophica produced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

Abstract: Provided is a method for selectively obtaining, for a given target gene, a “joined DNA fragment” wherein just a target gene fragment is joined with desired other DNA fragments, regardless of whether a DNA fragment containing a target gene sequence has been purified. In the provided method, a double-stranded joining DNA fragment containing a sequence A and/or a sequence B is selectively joined to the ends of a target gene fragment. A mixture of a double-stranded gene fragment, the 3? end of which is protruding, and the double-stranded joining DNA fragment, which are related in a prescribed manner, undergoes at least two cycles of thermal denaturation, reassociation, and DNA synthesis, resulting in a “joined DNA fragment,” which is a double-stranded DNA fragment including at least one instance of a sequence resulting from joining sequence A, the target gene sequence, and sequence B. A “single-side joined DNA fragment” can also be obtained, by a similar method.

Abstract: A method of correcting singletons in a selected AAV sequence in order to increasing the packaging yield, transduction efficiency, and/or gene transfer efficiency of the selected AAV is provided. This method involves altering one or more singletons in the parental AAV capsid to conform the singleton to the amino acid in the corresponding position(s) of the aligned functional AAV capsid sequences.

Abstract: The present invention relates to a Lyme disease vaccine, a genetic construct, recombinant protein, method for genetic construct design, method for vaccine delivery, method for recombinant proteins delivery, use of recombinant proteins in the production of Lyme disease vaccine. In particular, the method concerns the use of TROSPA and TROSPA-Salp15 recombinant proteins derived from castor bean tick (Ixodes riccinus) as a component of Lyme disease vaccine for animals. The antibodies present in blood of an immunized vertebrate directed against the TROSPA proteins considerably reduce the chance of infecting new ticks by blocking or hindering the interaction of TROSPA protein with OspA protein of Borrelia burgdorferi sensu lato. The interaction is crucial in the process of the spirochete entering a tick.

Abstract: The present invention is related to a nucleic acid construct comprising an expression unit for the expression of E1B, wherein the expression unit comprises a promoter, a nucleotide sequence coding for E1B, and a 3?UTR, wherein the promoter is operatively linked to the nucleotide sequence coding for E1B, wherein the 3?UTR comprises 30 or less than 30 Exonic Enhancer Elements (ESEs), preferably 20 or less than 20 Exonic Enhancer Elements (ESEs), and wherein the 3? UTR is a non-viral 3? UTR.

Abstract: The present application applies to the field of veterinary vaccines, in particular of vaccines for poultry against avian influenza. The vaccine is based on a recombinant viral vector expressing the haemagglutinin protein of an influenza virus, wherein the vector is herpes virus of turkeys (HVT) and the haemagglutinin gene is driven by a glycoprotein B gene promoter from a mammalian herpesvirus. A vaccine comprising this HVT+HA vector can be used to induce a protective immune response against avian influenza in poultry, and to reduce the spread of AIV. The invention also relates to methods, uses, and vaccines involving the HVT+HA vector.

Abstract: Circular nucleic acid vectors that provide for persistently high levels of protein expression are provided. The circular vectors of the subject invention are characterized by being devoid of expression-silencing bacterial sequences, where in many embodiments the subject vectors include a unidirectional site-specific recombination product hybrid sequence in addition to an expression cassette. Also provided are methods of using the subject vectors for introduction of a nucleic acid, e.g., an expression cassette, into a target cell, as well as preparations for use in practicing such methods. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications. Also provided is a highly efficient and readily scalable method for producing the vectors employed in the subject methods, as well as reagents and kits/systems for practicing the same.

Abstract: The present invention provides means and methods for simple and efficient introduction of foreign genetic material into the plant cell. Particularly, the present invention combines seed priming and virus-based DNA constructs for efficient introduction of heterologous DNA into plants.

Abstract: Materials and methods useful for generating highly mannosylated pseudotyped lentiviral vector particles comprising a Vpx protein are provided.

Abstract: The present invention relates to the construction and utilization of a new mammalian expression vector that contains a unique multiple cloning site (MCS), designated pUHAB. The pUHAB vector comprises a high copy replication origin (ColE1), a drug resistance gene (TK-Hygromycin), and a human cytomegalovirus promoter operably associated with a unique intron (hCMV/intron). Further, pUHAB comprises a selectable marker conferring resistance to kanamycin in bacterial cells, and a phage f1(+) region. pUHAB can be used to transiently or stably express cloned genes when transfected into mammalian cells. The invention also encompasses kits and host cells and cell lines comprising pUHAB, and methods of producing a recombinant protein using pUHAB.

Abstract: The present invention relates to the field of functional analysis of Jatropha genes on a genomic scale. More specifically, the present invention relates to a method for high-throughtput functional analysis of Jatropha curcas genes on a genomic scale using virus-induced gene silencing. The method involves use of the tobacco rattle virus (TRV).

Abstract: The present invention discloses a cell system, as a host cell to be infected with an F gene-deficient virus, which can constitutively and stably express the F protein, and a method for producing an F gene-deficient virus by utilizing the cell. A non-proliferative human parainfluenza type 2 virus vector is produced by co-culturing an F gene-deficient human parainfluenza type 2 virus with a Vero cell having the F gene of human parainfluenza type 2 virus in such a manner that the F gene is non-inducibly expressed, and isolating viral particles from a culture supernatant.

Abstract: A plasmid is provided comprising the following functional units: a prokaryotic origin of replication, a marker sequence, two specific recombinase recognition sequences and a multiple cloning site, whereby it comprises a gene coding for a sequence specific recombinase, whereby the units are arranged on the plasmid in such a way that the plasmid is divided into a miniplasmid and a minicircle upon expression of the sequence specific recombinase, said miniplasmid comprising the prokaryotic origin of replication, the marker sequence and the gene for the sequence specific recombinase and said minicircle comprising the multiple cloning site.

Abstract: The invention provides coding and promoter sequences for a VS-1 and AP-2 gene, which affects the developmental process of senescence in plants. Vectors, transgenic plants, seeds, and host cells comprising heterologous VS-1 and AP-2 genes are also provided. Additionally provided are methods of altering nutrient allocation and composition in a plant using the VS-1 and AP-2 genes.

Abstract: The present invention relates to an optimized baculovirus construct useful for the production of virus (-like) particles or viral vectors, in particular viral vectors for gene therapy.

Abstract: Materials and methods are provided for replacing one or more amino acids in a polypeptide with an amino acid of choice to form mutant proteins. Both naturally and non-naturally occurring amino acids can be inserted. A population of mutant proteins can be created in which an amino acid residue has replaced an existing residue at random locations along the primary sequence of the protein. The provided techniques allow for the study of proteins and development of proteins with improved functionalities.

Abstract: The present invention discloses a cell system, as a host cell to be infected with an F gene-deficient virus, which can constitutively and stably express the F protein, and a method for producing an F gene-deficient virus by utilizing the cell. A non-proliferative human parainfluenza type 2 virus vector is produced by co-culturing an F gene-deficient human parainfluenza type 2 virus with a Vero cell having the F gene of human parainfluenza type 2 virus in such a manner that the F gene is non-inducibly expressed, and isolating viral particles from a culture supernatant.

Abstract: The present provides fusion proteins comprising PDGF and VEGF binding portions, and recombinant viral particles encoding the fusion proteins. Compositions comprising the fusion proteins and viral particles as well as methods of using the same are also provided.

Abstract: This invention relates to methods, compositions, and kits for enzymatic phosphorothioation of the sugar-phosphate backbone of nucleic acids. The invention allows for phosphorothioation of pre-existing nucleic acids.

Abstract: A method of mRNA production for use in transfection is provided, that involves in vitro transcription of PCR generated templates. This RNA can efficiently transfect different kinds of cells. This approach results in increased efficiency (fidelity and productivity) of mRNA synthesis and is less time consuming because it does not require cloning, and also consequently eliminates the unwanted errors and effects related to RNA made on DNA templates obtained with cloning techniques. The results of transfection of RNAs demonstrate that RNA transfection can be very effective in cells that are exceedingly difficult to transfect efficiently with DNA constructs. The method can be used to deliver genes into cells not- or only poorly transfectable for DNA, in vitro and in vivo.

Abstract: The present disclosure describes nucleic acids, and viruses comprising such nucleic acids, for growing a toxic gene in an insect cell. These nucleic acids comprise a sequence encoding a toxic polypeptide, and an intron that interrupts the sequence, whereby the intron is spliced in mammalian cells but not in insect cells. Infection of mammalian cells but not insect cells with the nucleic acids or viruses can lead to expression of toxic levels of the toxic polypeptide in mammalian cells but not in insect cells. Viruses, such as an AAV or a baculovirus comprising a nucleic acid can be grown in insect cell lines in vitro and can be administered to a subject in need of therapy, such as a subject in need of cancer therapy.

Abstract: The present invention generally relates to methods and compositions used delivery of gene editing compositions including transcriptional effectors with parvovirus and preferred methods for making same.

Abstract: The present invention describes a recombinant retroviral vector which cannot by itself achieve complete reverse transcription as well as its uses, in particular for transiently transferring in vitro, ex vivo or in vivo at least one ribonucleic acid sequence of interest in a cell. Such a transient transgene expression is of interest in the context of research, therapy and more generally in the field of biotechnology.

Abstract: The invention relates to a nucleic acid preparation with a content of below 1% protein, preferably below 0.1% protein, free of ethidium bromide, phenol, cesium chloride and detergents based on octyl phenol polyethylene glycol ether)n and with a content of below 1 EU/mg DNA of endotoxins. Said preparation is suitable as a drug particularly in gene therapy.

Abstract: The present disclosure provides non-naturally occurring miR-196a-2 miRNAs and non-naturally occurring miR-204 miRNAs. The non-naturally occurring miRNAs of the disclosure have mature strand sequences distinct from their endogenous counterparts. The disclosure also provides methods of selecting mature strand sequences that function optimally in non-naturally occurring miR-196a-2 miRNAs. The methods and compositions of the disclosure may be used to mediate gene silencing via the RNAi pathway.

Abstract: Compositions and methods of treatments of cells are provided for altering the phenotype of a cell by administering an oligonucleotide complex to the cell, the complex having two strands and chemical modifications.

Abstract: The present invention provides synthetic 5?UTRs comprising a first polynucleotide fragment and a second polynucleotide fragment, wherein the first polynucleotide fragment comprises at least one splice site of a first eukaryotic gene, the second polynucleotide fragment comprises at least a portion of 5? untranslated region of a second eukaryotic gene, and the first polynucleotide fragment is located 5? of the second polynucleotide fragment. In one embodiment, the first polynucleotide fragment comprises the second intron of a sarcoplasmic/endoplasmic reticulum calcium ATPase gene and the second polynucleotide fragment comprises at least a portion of the 5? untranslated region (5?UTR) of a eukaryotic casein gene. The synthetic 5?UTRs are useful for increasing the expression of a transgene when positioned between a promoter and a transgene within an expression vector.

Abstract: This invention provides transcription regulatory control sequences, the activity of which function as biomarkers for a variety of biological responses. This invention also provides expression constructs in which a biomarker transcription regulatory sequence is operably linked with a sequence for a reporter. Cells that comprise these expression constructs can be used in assays to identify conditions that modulate activity of the biological response.

Abstract: Cell lines are provided for expressing peptidylglycine alpha-amidating monooxygenase (PAM), or one of its two catalytic domains. High levels of enzyme expression are achieved while utilizing a non-animal source, low protein tissue culture medium. A robust two-step downstream purification results in high enzyme purity. Resulting PAM, or its PHM catalytic domain, is used to catalyze the enzymatic conversion of X-Gly to X-alpha-hydroxy-Gly or X—NH2 (X being a peptide or any chemical compound having a carbonyl group to which a glycine group can be covalently attached). Methods of preparing preferred cell lines are also set forth.

Abstract: Described is a chimerical adenovirus-parvovirus vector characterized in that it comprises the entire parvovirus genome inserted into a adenovirus genome, wherein: (a) the adenovirus genome is characterized by deletion of E1 and E3, and (b) the biological activity of the parvoviral protein NS, preferably NS1, is reduced or eliminated.

Abstract: The present invention is directed to Herpes simplex-2 viruses that may be used in vaccines to immunize patients against genital herpes.