Abstract: Based on the three-dimensional structure of albumin, the inventors have designed variant polypeptides (muteins) which have one or more cysteine residues with a free thiol group (hereinafter referred to as “thio-albumin”). The variant polypeptide may be conjugated through the sulphur atom of the cysteine residue to a conjugation partner such as a bioactive compound.

Abstract: Some embodiments of the present invention generally relate to the novel method, compositions and applications of mechanosensitive channels in therapeutics for visual, neurological and other disorders. Specifically, some embodiments of the invention relate to exploitation of mechanosensitive channels' intrinsic property as a transgenic pressure modulator and alternative outflow actuator in the treatment of different diseases such as glaucoma, dry eye disease and other diseases involving of cells prone to mechanical or osmotic stress. Furthermore, some embodiments of the invention relate to exploitation of mechanosensitive channels for stimulation of cells as well as molecular delivery to cells triggered by internal as well as external mechanical stimuli.

Abstract: Methods of expressing an expression product of interest are provided. Accordingly there is provided a method comprising introducing into a cell a polynucleotide comprising an AimR responsive element operatively linked to a nucleic acid sequence encoding the expression product of interest, and contacting said cell with an AimP peptide comprising an amino acid sequence of XXXXGG/A, wherein said AimP peptide is capable of binding said AimR polypeptide and dissociating said AimR polypeptide from said AimR responsive element. Also provided are articles of manufacture, isolated peptides, polynucleotides and nucleic acid constructs.

Abstract: The present invention relates to microfluidic fluidic systems and methods for the in vitro modeling diseases of the lung and small airway. In one embodiment, the invention relates to a system for testing responses of a microfluidic Small Airway-on-Chip infected with one or more infectious agents (e.g. respiratory viruses) as a model of respiratory disease exacerbation (e.g. asthma exacerbation). In one embodiment, this disease model on a microfluidic chip allows for a) the testing of anti-inflammatory and/or anti-viral compounds introduced into the system, as well as b) the monitoring of the participation, recruitment and/or movement of immune cells, including the transmigration of cells. In particular, this system provides, in one embodiment, an in-vitro platform for modeling severe asthma as “Severe Asthma-on-Chip.” In some embodiments, this invention provides a model of viral-induced asthma in humans for use in identifying potentially effective treatments.

Abstract: The invention relates to, in part, improved methods for the production of beta-lactamase using Escherichia coli (E. coli) cells. High yield production of beta-lactamase is achieved using methods of the invention.

Abstract: The invention is directed to antibacterial compositions comprising bacteria modified to comprise phasmids engineered to deliver of CRISPR RNAs and methods for their use.

Abstract: Lipids can be extracted from a microbial biomass that constitutes at least 20% lipids by weight and has a moisture content of less than 4% by weight by applying pressure to the biomass so as to release lipids therefrom, thereby leaving a biomass of reduced lipid content; and collecting the lipids.

Abstract: The present invention relates to mannanase variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: Systems and methods for modeling a three-dimensional protein structure are disclosed. The method includes receiving a primary amino acid sequence of a three-dimensional protein, translating the primary amino acid sequence to a first vector, determining a per-residue conformation index for each amino acid residue in the primary amino acid sequence, determining a vector set for each amino acid residue in the primary amino acid sequence, and using the per-residue interaction vector set to generate a multi-dimensional matrix for the three-dimensional protein structure. The first vector includes a unique numerical descriptor value corresponding to each amino acid residue in the primary amino acid sequence. The vector set includes a plurality per-residue interaction factors corresponding to a plurality of conformation indexes for that amino acid residue.

Abstract: A mutant SaCas9 protein such as a protein having an amino acid sequence resulting from mutations of glutamic acid at the 782-position to lysine (E782K), leucine at the 800-position to arginine (L800R), asparagine at the 968-position to arginine (N968R), asparagine at the 985-position to alanine (N985A), arginine at the 991-position to alanine (R991A), alanine at the 1021-position to serine (A1021S), threonine at the 927-position to lysine (T927K), lysine at the 929-position to asparagine (K929N), and isoleucine at the 1017-position to phenylalanine (I1017F) in SEQ ID NO: 2 has relaxed restriction on target sequence while maintaining binding ability to guide RNA, and is useful as a tool for gene editing.

Abstract: The disclosure provides systems, methods, and compositions for a target specific nuclease and a blunting enzyme to correct frameshift mutations for genome editing and treatment of diseases. In some embodiments, the target specific nuclease and the blunting enzyme are combined with a guide RNA and/or a microhomology-mediated end joining (MMEJ) inhibitor.

Abstract: The invention relates to modified helicases with reduced unbinding from polynucleotides. The helicases can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.

Abstract: The invention relates to a new method of characterising a target polynucleotide. The method uses a pore and a Dda helicase. The helicase controls the movement of the target polynucleotide through the pore. The invention also relates to modified Dda helicases which can be used to control the movement of polynucleotides and are particularly useful for sequencing polynucleotides.

Abstract: The invention is in the field of regulation of enzymatic activity in nucleic acid modifying reactions. It describes a method of regulating enzymatic activity by adding chelating agents to the reaction composition and exploits the fact that both the binding of divalent cations to these chelating agents and the pH of commonly used buffers is temperature dependent. PCR experiments that are hampered by non-specific side products can be regulated such that the target sequence is amplified in a more specific manner.

Abstract: The disclosure provides a guide RNA (gRNA) comprising a DNA-binding domain and a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease protein-binding domain, wherein the DNA-binding domain is complementary to a target domain from a variant NRF2 gene found in a cancer cell but not in a non-cancerous cell. The disclosure also provides nucleic acid sequence encoding the gRNA. The disclosure further provides a method of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease and a guide RNA that is complementary to a target domain from a variant NRF2 gene in the subject.

Abstract: The present invention is directed to methods and compositions for typing of Lactobacillus buchneri bacterial strains, detecting the presence of a L. buchneri in a sample, identifying a strain of L. buchneri having resistance to an invasive foreign genetic element, modifying the resistance of bacteria and archeae to an invasive foreign genetic element, and introducing nicks into or cleaving double stranded DNA for genome editing.

Abstract: Methods and compositions for making bacteriocins are described in some embodiments herein. In some embodiments, pro-polypeptide comprising the bacteriocins in the desired ratios in cis, and separated by cleavage sited can be produced by a microbial cell comprising a nucleic acid encoding the pro-polypeptide. In some embodiments microfluidic devices and methods for making specified mixtures of antimicrobial peptides and/or bacteriocins are described.

Abstract: A protein expression system for use in a prokaryotic host is provided, the expression system comprising: a) an expression cassette comprising a nucleic acid sequence encoding a protein of interest operably linked to a T7 RNA polymerase-dependent promoter; and b) an expression cassette comprising a nucleic acid sequence encoding T7 RNA polymerase operably linked to a host polymerase-dependent ? phage promoter and a single perfect palindrome operator sequence; wherein the expression cassette for T7 RNA polymerase is located on the chromosome of a host cell.

Abstract: Epigenomic states of genome DNA are altered at multiple sites to rapidly change traits by providing a fusion protein including a first region that defines a polypeptide capable of binding sequence-specifically to multiple sites on genome DNA and a second region that defines a polypeptide capable of regulating an epigenomic state.

Abstract: This disclosure provides compositions, recombinant expression constructs, and engineered systems that include a polynucleotide including or encoding a Cas12a tracrRNA, and methods for their use. The materials and methods of the disclosure are especially suited to sequence-specific genome editing of eukaryotic genomic sequences.