Abstract: The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.

Abstract: The disclosure provides for compositions, methods and kits, for reducing off-target effects of genome engineering.

Abstract: The present disclosure provides engineered cross-type-nucleic-acid targeting nucleic acids and compositions thereof. Nucleic acid sequences encoding the engineered cross-type-nucleic-acid targeting nucleic acids, as well as expression cassettes, vectors and cells comprising such nucleic acid sequences, are described. Also, methods are disclosed for making and using the engineered cross-type-nucleic-acid targeting nucleic acids and compositions thereof.

Abstract: The present disclosure provides compositions and methods for enhancing directed nucleic acid repair, which are useful in the area of genome engineering.

Abstract: The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.

Abstract: The present specification discloses engineered Type II CRISPR-Cas9 systems comprising split-nexus Cas9-associated polynucleotides (sn-casPNs), including systems comprising three split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN/sn3-casPN) and systems comprising two split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN). Together with a Cas9 protein, the sn-casPNs facilitate site-specific modifications, including cleavage and mutagenesis, of a target polynucleotide in vitro and in vivo. Furthermore, the engineered Type II CRISPR-Cas9 systems comprising sn-casPNs are useful in methods of regulating expression of a target nucleic acid. Methods are described herein for the creation of a variety of engineered Type II CRISPR-Cas9 systems comprising two or more sn-casPNs. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.

Abstract: The present specification discloses engineered Type II CRISPR-Cas9 systems comprising split-nexus Cas9-associated polynucleotides (sn-casPNs), including systems comprising three split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN/sn3-casPN) and systems comprising two split-nexus Cas9-associated polynucleotides (sn1-casPN/5n2-casPN). Together with a Cas9 protein, the sn-casPNs facilitate site-specific modifications, including cleavage and mutagenesis, of a target polynucleotide in vitro and in vivo. Furthermore, the engineered Type II CRISPR-Cas9 systems comprising sn-casPNs are useful in methods of regulating expression of a target nucleic acid. Methods are described herein for the creation of a variety of engineered Type II CRISPR-Cas9 systems comprising two or more sn-casPNs. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described.

Abstract: The present specification discloses engineered Type II CRISPR-Cas9 systems comprising split-nexus Cas9-associated polynucleotides (sn-casPNs), including systems comprising three split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN/sn3-casPN) and systems comprising two split-nexus Cas9-associated polynucleotides (sn1-casPN/sn2-casPN). Together with a Cas9 protein, the sn-casPNs facilitate site-specific modifications, including cleavage and mutagenesis, of a target polynucleotide in vitro and in vivo. Furthermore, the engineered Type II CRISPR-Cas9 systems comprising sn-casPNs are useful in methods of regulating expression of a target nucleic acid. Methods are described herein for the creation of a variety of engineered Type II CRISPR-Cas9 systems comprising two or more sn-casPNs. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described.

Abstract: Systems, apparatuses, and methods are provided for developing an improved route from a navigation mesh. A route is provided, wherein the route includes a plurality of connected nodes having a starting node, an ending node, and internal nodes. Each internal node of the plurality of connected nodes is scored by an algorithm, wherein the score provided for each internal node based on an importance of the respective internal node to the overall shape of the route. The internal node with a score having a least importance on the shape of the route is removed, so long a valid path between the starting node and the ending node is maintained. The process may be repeated until a threshold simplification of the route has been achieved or until no further internal nodes can be removed without creating an invalid path.

Abstract: Class 2 CRISPR-Cas nucleoprotein complexes are disclosed comprising a Class 2 CRISPR-Cas protein, a CRISPR-Cas associated polynucleotide lacking a spacer element (casPN), and a distinct spacer element sequence polynucleotide (sesPN) comprising a target nucleic acid binding sequence. These complexes are capable of site-directed binding to a target nucleic acid complementary to the target nucleic acid binding sequence of the sesPN. The Class 2 CRISPR-Cas nucleoprotein complexes facilitate site-specific modifications, including cleavage and mutagenesis, of a target nucleic acid sequence. Polynucleotide sequences, expression cassettes, vectors, compositions, and kits for carrying out a variety of methods are also described.

Abstract: The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

Abstract: Systems, apparatuses, and methods are provided for developing an improved route from a navigation mesh. A route is provided, wherein the route includes a plurality of connected nodes having a starting node, an ending node, and internal nodes. Each internal node of the plurality of connected nodes is scored by an algorithm, wherein the score provided for each internal node based on an importance of the respective internal node to the overall shape of the route. The internal node with a score having a least importance on the shape of the route is removed, so long a valid path between the starting node and the ending node is maintained. The process may be repeated until a threshold simplification of the route has been achieved or until no further internal nodes can be removed without creating an invalid path.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof.

Abstract: The present disclosure provides DNA-guided CRISPR systems; polynucleotides comprising DNA, RNA and mixtures thereof for use with CRISPR systems; and methods of use involving such polynucleotides and DNA-guided CRISPR systems.

Abstract: This disclosure provides for compositions and methods for the use of nucleic acid-targeting nucleic acids and complexes thereof. Genome engineering can refer to altering the genome by deleting, inserting, mutating, or substituting specific nucleic acid sequences. The altering can be gene or location specific. Genome engineering can use nucleases to cut a nucleic acid thereby generating a site for the alteration. Engineering of non-genomic nucleic acid is also contemplated.

Abstract: A hard link, which is connected to a moldboard of a machine, includes a first range of float capability and a second range of float capability. The second range of float capability may provide a greater float height to the moldboard than the first range of float capability. In another embodiment, a machine includes a moldboard, a jib arm operatively configured to raise and lower the moldboard, and a link attaching the jib arm to the moldboard. The link includes two discrete float capabilities.