Abstract: The present invention relates to modified guide RNAs and their use in clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems.

Abstract: Vectors expressing kanA-kanB-kanK and other kanamycin production-related genes, Streptomyces species recombinant bacteria transformed with the vectors, a method of producing kanamycin antibiotics by the bacteria, and a new kanamycin compound produced by the bacterium are provided. With the use of the recombinant bacteria of the present invention, the direct fermentative biosynthesis of amikacin and tobramycin as semi-synthetic kanamycins is possible, and the yield of kanamycin B as a precursor of the semi-synthetic kanamycin is improved.

Abstract: This disclosure relates to a method of generating conditionally active biologic proteins from wild type proteins, in particular therapeutic proteins, which are reversibly or irreversibly inactivated at some physiological conditions. For example, conditionally active biologic proteins are active in tumors, but virtually inactive at other body parts, or conditionally active antibodies cap able of crossing blood-brain-barrier.

Abstract: This invention provides high growth capacity strains of auxotrophic Escherichia coli and methods for generating thereof. The high growth capacity strains express a complementing auxotrophic plasmid that allows the strain to grow in the absence of the auxotrophic amino acid. Also, provided herein is a method for preparing a bacterial cell extract of a high growth capacity strain of auxotrophic Escherichia coli for use in an in vitro protein expression.

Abstract: The present invention relates to an interleukin-2 expression construct for yeast, comprising a methanol oxidase (MOX) promoter; a human serum albumin gene or a fragment thereof; and an interleukin-2 (IL-2) gene, and to a yeast comprising the expression construct. The interleukin-2 expression construct for yeast according to the present invention makes it possible to produce an expressed and secreted fusion protein of human serum albumin (HSA) and interleukin-2 at low costs and easily separate recombinant interleukin-2 from the fusion protein. Thus, the interleukin-2 expression construct for yeast may be effectively used to produce a large amount of recombinant interleukin-2 with high purity.

Abstract: The invention includes materials and methods to generate numerous small RNAs from one polynucleotide construct (synthetic gene) to facilitate RNA-guided multiplex genome editing, modification, inhibition of expression and other RNA-based technologies. The synthetic gene/polynucleotide construct encodes polycistronic RNA components separated by tRNAs, and preferably also includes regulatory components such as a promoter or terminator to form an expression cassette. Once transcribed in a cell, the transcript is processed by the cell to multiple RNA molecules by the endogenous tRNA processing system. The system can be used for any RNA based gene manipulation method including RNA-mediated genome editing, artificial microRNA mediated gene silencing, small RNA mediated genetic manipulation, double-stranded RNA mediated gene silencing, antisense mechanisms and the like.

Abstract: The present invention relates to the field of fungal biotechnology, more particularly to genetic engineering methods for the production of carotenoids in fungal hosts selected from Rhodospordium and Rhodotorula genera.

Abstract: Described herein are methods for predicting a benefit from inclusion of taxane in a chemotherapy regimen in a patient suffering from or at risk of developing recurrent neoplastic disease, in particular breast cancer. Said method includes the steps of determining the expression levels of the marker genes S100P and PCSK6 and mathematically combining the expression level values to yield a combined score and comparing said combined score to a reference-value, wherein a high combined score is indicative of a benefit from including a taxane in a chemotherapy regimen of said patient and a low combined score is indicative of not having a benefit from including a taxane in a chemotherapy regimen.

Abstract: Methods and compositions are provided for modifying one or more target loci in a cell. Such methods comprise providing a cell comprising a first polynucleotide encoding a first selection marker operably linked to a first promoter active in the cell, wherein the first polynucleotide further comprises a first recognition site for a first nuclease agent. A first nuclease agent is introduced into a cell, wherein the first nuclease agent induces a nick or double-strand break at the first recognition site. Further introduced into the cell is a first targeting vector comprising a first insert polynucleotide flanked by a first and a second homology arm that correspond to a first and a second target site located in sufficient proximity to the first recognition site. At least one cell is then identified comprising in its genome the first insert polynucleotide integrated at the target locus.

Abstract: A microorganism which is genetically modified so that it produces a first essential biomass precursor by metabolizing CO2 using a recombinant carbon fixation enzyme is disclosed. The microorganism produces a second biomass precursor by metabolizing an organic carbon source and not by metabolizing CO2. The microorganism does not use the organic carbon source for producing the first essential biomass precursor.

Abstract: The present invention provides compositions and methods for the production and delivery of recombinant double-stranded RNA molecules (dsRNA) encoding heterologous proteins, which can be useful for various therapeutic purposes as well as for the production of desired proteins. The compositions contain engineered double-stranded RNA particles (dsRPs) that can contain a double-stranded RNA molecule that can be a genome or portion of a genome, which can be enclosed in a capsid or coat protein. The dsRNA molecule also comprises an RNA sub-sequence that encodes a heterologous protein. The dsRPs can be derived from wild-type viral organisms. The delivery of the dsRPs (or DNA or RNA molecules) of the invention to an organism provides for therapeutic benefits as well as for the production of desired proteins.

Abstract: Biomarkers, methods, assays, and kits are provided for predicting the efficacy of adjuvant chemotherapy (ACT) in a subject with early-stage non-small cell lung cancer (NSCLC).

Abstract: The present disclosure provides methods for processing or analyzing a sample of tissue of a subject, to generate a classification of the sample of tissue as positive or negative for thyroid cancer. The present disclosure also provides algorithms and methods of classifying cancer, for example, thyroid cancer, methods of determining molecular profiles, and methods of analyzing results.

Abstract: Compositions, methods and kits are disclosed for high-sensitivity single molecule digital counting by the stochastic labeling of a collection of identical molecules by attachment of a diverse set of labels. Each copy of a molecule randomly chooses from a non-depleting reservoir of diverse labels. Detection may be by a variety of methods including hybridization based or sequencing. Molecules that would otherwise be identical in information content can be labeled to create a separately detectable product that is unique or approximately unique in a collection. This stochastic transformation relaxes the problem of counting molecules from one of locating and identifying identical molecules to a series of binary digital questions detecting whether preprogrammed labels are present. The methods may be used, for example, to estimate the number of separate molecules of a given type or types within a sample.

Abstract: The present invention relates to a method for fine-tuning gene expression levels using a synthetic regulatory sRNA in a prokaryotic cell. The present invention can simultaneously, easily, and quickly apply various target gene combinations to various strains without gene deletion through the synthetic regulatory sRNA for regulating gene expression and is therefore very suitable for measuring the metabolizability of each strain and selecting an optimum strain. In addition, the method has the advantages of easily and quickly selecting a target gene for the inhibition of gene expression and expressing the gene thus selected to a desired degree and thus can be used in producing recombinant strains for the efficient production of various metabolites and establishing a method for the efficient production and is therefore very useful.

Abstract: Provided is an optically controlled gene expression system of prokaryotic bacterium, comprising: a) a photosensitive recombinant transcription factor encoding gene, the photosensitive recombinant transcription factor is one fusion protein comprising a first polypeptide as the DNA bonding domain and a second polypeptide as the photosensitive domain; b) a target transcription unit comprising promoter or promoter-reaction element or reaction element-promoter containing at least one reaction element recognized/bound by the first polypeptide and the nucleic acid sequence to be transcribed. Also provided is a prokaryotic expression vector comprising said optically controlled gene expression system, and a method for regulating gene expression in a prokaryotic host cell by using the optically controlled gene expression system. Also provided is a reagent kit containing different components of the optically controlled gene expression system.

Abstract: Algorithm-based molecular assays that involve measurement of expression levels of prognostic and/or predictive genes, or co-expressed genes thereof, from a biological sample obtained from a cancer patient, and analysis of the measured expression levels to provide information concerning the likelihood of recurrence of colorectal cancer and/or the likelihood of a beneficial response to chemotherapy for the patient are provided herein. Methods of analysis of gene expression values of prognostic and/or predictive genes, as well as methods of identifying gene expression-tumor region ratios, tumor-associated stromal surface area, and gene cliques, i.e. genes that co-express with a validated biomarker and thus may be substituted for that biomarker in an assay, are also provided.

Abstract: The present invention provides compositions and methods for the production and delivery of recombinant double-stranded RNA molecules (dsRNA) targeting pathogen sequences, which can be useful as an RNA vaccine. The compositions contain engineered double-stranded RNA particles (dsRPs) that can contain a double-stranded RNA molecule that can be a genome or portion of a genome, which can be enclosed in a capsid or coat protein. The dsRNA molecule also comprises an RNA sub-sequence that binds to a target sequence of a pathogenic organism. The dsRPs can be derived from wild-type viral organisms. The delivery of the dsRPs of the invention to an organism provides a protection to the organism from the pathogen.

Abstract: The present invention relates to a method for preparing a recombinant Phe-free or Phe-low protein, the method comprising using B. subtilis or B. licheniformis as an expression system and/or a recombinant host cell into which a nucleotide encoding a recombinant Phe-free or Phe-low protein has been inserted into the genome.

Abstract: The present invention relates to a nucleic acid sequence, comprising or coding for a coding region, encoding at least one peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, at least one histone stem-loop and a poly(A) sequence or a polyadenylation signal. Furthermore the present invention provides the use of the nucleic acid for increasing the expression of said encoded peptide or protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine, e.g. for use in the treatment of infectious diseases. The present invention further describes a method for increasing the expression of a peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal.