9 results about "DNA Endonuclease" patented technology

The invention relates to circular DNA molecule for rescuing recombinant adenoviruses comprising a recombinant adenoviral genome with two inverted terminal repeats (ITRs) flanking the genome ends, wherein at least one of the ITRs is associated with a target sequence adjacent to a PAM sequence, wherein the target sequence is configured for generating an RNA-guided DNA endonuclease-mediated DNA double strand break at the external end of or in close proximity outside the external end of the respective ITR, preferably within less than about 15 nucleotides. The invention also relates to a kit and a method for rescuing recombinant adenoviruses comprising or using a circular DNA molecule as described herein.

Provided herein are programmable RNA-guided DNA endonucleases and derivatives and uses thereof and methods of using the same.

The present disclosure provides novel mirror image CRISPR compositions that include components such as mirror image CRISPR-associated endonucleases and mirror image guide RNA for processing of mirror image nucleic acids. In some embodiments, the composition is a mirror image DNA-cleaving and / or modifying composition including a D-form DNA endonuclease (e.g., a D-form Cas protein) and a L-form guide RNA that includes a sequence complementary to a target sequence of a target L-DNA. In some embodiments, the composition further comprises a D-form DNA ligase (e.g., a D-form T4 ligase protein). Also provided are D-form Cas protein and D-form T4 ligase protein compositions, synthetic precursors thereof, and methods of preparing the same.

The application discloses a method for rapidly and non-traceably preparing mRNA containing long polyadenine and application thereof. Through modular design and an IIS type restriction DNA endonuclease system, complex gene synthesis containing long poly(A) is decomposed into two steps of rapid synthesis of simple target genes and non-traceable connection with a previously prepared leading plasmid. The mRNA synthesized based on the method can carry a fixed length of poly(A) without tailing operation, and since the target gene synthesis does not involve a long poly(A) complex fragment, the speed and efficiency of the template plasmid preparation are greatly improved. The plasmid can be directly used as a linearized template for in vitro transcription synthesis of long polyadenosine deoxyribonucleic acid poly(A) mRNA, the IIS type restriction DNA endonuclease system does not need to introduce an additional splicing sequence between the target gene and poly(A), and the transcribed mRNA sequence completely meets the expectation.

Isolation or in vitro assembly of the Cas9-crRNA complex of the Streptococcus thermophilus CRISPR3 / Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule using an RNA-guided DNA endonuclease comprising at least one RNA sequence and at least one of an RuvC active site motif and an HNH active site motif; for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one point mutation; for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro; and for re-programming a Cas9-crRNA complex specificity in vitro or using a cassette containing a single repeat-spacer-repeat unit.

The invention provides a method for selectively accumulating genetic modifications in an offspring cell of a microbial species, wherein the microbial species has (i) a first mating type and (ii) a second mating type compatible with the first mating type; and wherein the method comprises a mating cycle comprising: a preparation stage comprising providing a first cell and a second cell of the microbial species, the first cell having the first mating type and the second cell having the second mating type, wherein the first cell and the second cell have a first locus, a second locus, a third locus, a fourth locus, and a fifth locus, wherein preferably at least the fourth locus and the fifth locus are chromosomal loci, and wherein: the first locus of the first cell encodes a first guide RNA, wherein the first guide RNA is configured to hybridize with a first target sequence, wherein the first locus of the second cell comprises the first target sequence; the second locus of the second cell encodes a second guide RNA, wherein the second guide RNA is configured to hybridize with a second target sequence, wherein the second locus of the first cell comprises the second target sequence; the third locus of the first cell and the third locus of the second cell encode an RNA-guided DNA endonuclease, wherein the RNA-guided DNA endonuclease is configured to functionally associate with the first guide RNA and with the second guide RNA; the fourth locus of the first cell comprises a first mating-type promoter configured to facilitate expression of a first selectable marker providing a first selectable trait in cells having the first mating type; the fourth locus of the second cell comprises a second mating-type promoter configured to facilitate expression of a second selectable marker providing a second selectable trait in cells having the second mating type; the fifth locus of the first cell comprises a third selectable marker providing a third selectable trait; the fifth locus of the second cell comprises a fourth selectable marker providing a fourth selectable trait; and preferably the first selectable trait, the second selectable trait, the third selectable trait and the fourth selectable trait differ from each other; a mating stage comprising bringing the first cell and the second cell in contact with each other to provide an offspring cell population; and a selection stage comprising selecting the offspring cell from the offspring cell population or from cells derived therefrom, wherein the offspring cell is selected based on at least two selectable traits, wherein preferably the at least two selectable traits comprise one or more of (a) at least the third selectable trait and the fourth selectable trait, (b) at least the first selectable trait and the fourth selectable trait, and (c) at least the second selectable trait and the third selectable trait.

Provided herein are guide nucleic acid-guided DNA endonucleases and derivatives and uses thereof and methods of using the same.

The present disclosure provides novel mirror image CRISPR compositions that include components such as mirror image CRISPR-associated endonucleases and mirror image guide RNA for processing of mirror image nucleic acids. In some embodiments, the composition is a mirror image DNA-cleaving and / or modifying composition including a D-form DNA endonuclease (e.g., a D-form Cas protein) and a L-form guide RNA that includes a sequence complementary to a target sequence of a target L-DNA. In some embodiments, the composition further comprises a D-form DNA ligase (e.g., a D-form T4 ligase protein). Also provided are D-form Cas protein and D-form T4 ligase protein compositions, synthetic precursors thereof, and methods of preparing the same.

Provided is a composition or system. The composition or system comprises: a DNA endonuclease or a nucleic acid encoding the DNA endonuclease, a single-molecule DNA-targeting RNA or a DNA polynucleotide encoding the single-molecule DNA-targeting RNA; and a donor template comprising a nucleic acid sequence encoding the FIX protein or a variant or functional derivative thereof. The single-molecule DNA-targeting RNA is a transferrin gene-targeting RNA. When used for treating hemophilia, the provided composition or system can increase the expression of the FIX protein and prolong the activity of the FIX protein in the blood, thereby improving the therapeutic effect, reducing the patient's dependence on the treatment, and enhancing the stability and efficacy of long-term treatment.