39 results about "Meganuclease" patented technology

The present disclosure encompasses a lipid nanoparticle (LNP) comprising a polypeptide comprising a nucleic acid sequence encoding an engineered meganuclease that binds and cleaves a recognition sequence within a Hepatitis B virus (HBV) genome. Further, the disclosure encompasses pharmaceutical compositions comprising the LNPs, and the use of such compositions for inactivating a pol gene of an HBV genome or an HBV genome fragment in a cell and treating HBV infections or diseases associated with HBV infections.

To provide an engineered meganuclease that recognizes and cleaves the recognition sequence in the human T cell receptor alpha constant region gene.SOLUTION: Provided herein is an engineered meganuclease comprising a first subunit and a second subunit, where the first subunit binds to a first recognition half site of a specific sequence in human T cell receptor alpha constant region gene and comprises a first hypervariable region, and the second subunit binds to a second recognition half site of the specific sequence comprising the specific sequence and comprises a second hypervariable region, where the second hypervariable region has (a) at least 81% sequence homology to an amino acid sequence corresponding to residues 24 to 79 of a specific sequence or (b) at least 86% sequence homology to an amino acid sequence corresponding to residues 24 to 79 of a specific sequence.SELECTED DRAWING: None

Described herein are compositions and methods for reducing or eliminating latent herpes simplex virus type 2 (HSV-2) in HSV-2 infected cells, or for reducing or eliminating latent HSV-2 reactivation in HSV-2 infected cells, thereby providing viable therapeutic methods for latent HSV-2 infection. The compositions comprise a plurality of viral vectors, where the viral vectors comprise a sequence encoding a HSV-2 specific megasuclease. The method comprises delivering a plurality of one or more viral vectors to the HSV-2 infected cell, where each of the one or more viral vectors comprises a sequence encoding an HSV-2 specific megasuclease.

Disclosed herein are engineered meganucleases engineered to recognize and cleave a recognition sequence present in human mitochondrial DNA (mtDNA). The disclosure further relates to the use of such engineered meganucleases in methods for producing genetically-modified eukaryotic cells, and to a population of genetically-modified eukaryotic cells, wherein mutant mtDNA is modified or degraded.

Disclosed are engineered meganucleases that bind and cleave a recognition sequence within a serine peptidase inhibitor, Clade A, Member 1 (SERPINA1) gene, which encodes alpha-1 antitrypsin (AAT). Further disclosed are donor polynucleotides that encode functional AAT proteins. The present disclosure also encompasses methods of using such engineered meganucleases and donor polynucleotides to make genetically-modified cells and use of such compositions for treatment of AAT deficiency.

The present invention provides a genetically modified composition for treating cancer, and a method for preparing and using a genetically modified composition in the treatment of cancer. [Solution] The compositions and methods disclosed herein can be used to identify cancer-specific T cell receptors (TCRs) that recognize unique immunogenic mutations in a patient's cancer and to treat any type of cancer in the patient. Insertion of these transgenes encoding cancer-specific TCRs into T cells, using non-viral methods (e.g., CRISPR, TALEN, transposon-based ZEN, meganuclease, or Mega-TAL), is a novel technique that opens up new opportunities to extend immunotherapy to many cancer types.

Disclosed are recombinant meganucleases engineered to bind and cleave a recognition sequence present in a mutant RHO P23H allele. The invention further relates to the use of such recombinant meganucleases in a method for treating retinitis pigmentosa, wherein the mutant RHO P23H allele is preferentially targeted, cleaved, and inactivated.

The present invention includes engineered nuclease enzymes that recognize and cleave a recognition sequence within a first exon of a human T cell receptor (TCR) alpha constant region gene. The engineered broad-range nuclease may exhibit at least one optimized property, such as enhanced (i.e., increased) specificity or cleavage efficiency, as compared to the first generation broad-range nuclease TRC 1-2x. 87 EE. The invention also encompasses methods of making genetically modified cells using such engineered nuclease, as well as the use of such cells in pharmaceutical compositions and in methods of treating diseases such as cancer.

This disclosure covers engineered macronucleases that bind to and cleave recognition sequences within the dystrophin gene. This disclosure also covers methods for preparing genetically modified cells using such engineered macronucleases. Furthermore, this disclosure covers pharmaceutical compositions comprising engineered macronuclease proteins or polynucleotides encoding engineered macronucleases of this disclosure, and the use of such compositions for modifying the dystrophin gene in a subject or for treating Duchenne muscular dystrophy.

Rationally-designed LAGLIDADG meganucleases and methods of making such meganucleases are provided. In addition, methods are provided for using the meganucleases to generate recombinant cells and organisms having a desired DNA sequence inserted into a limited number of loci within the genome, as well as methods of gene therapy, for treatment of pathogenic infections, and for in vitro applications in diagnostics and research.

To provide means for creating T cells from a third party donor by reducing the expression of the endogenous T cell receptor and not initiating GVHD upon administration.SOLUTION: Disclosed herein are recombinant meganucleases engineered to recognize and cleave a recognition sequence present in the human T cell receptor alpha constant region gene. The present disclosure further relates to the use of such recombinant meganucleases in methods for producing genetically modified eukaryotic cells.SELECTED DRAWING: Figure 1

The present invention encompasses engineered meganucleases that bind and cleave a recognition sequence within the first exon of the human T cell receptor (TCR) alpha constant region gene. The engineered meganucleases can exhibit at least one optimized characteristic, such as increased specificity or efficiency of cleavage, when compared to previous generations of meganucleases. The present invention also encompasses methods of using such engineered nucleases to make genetically-modified cells, and the use of such cells in a pharmaceutical composition and in methods for treating diseases, such as cancer.

The present invention encompasses engineered nucleases which recognize and cleave a recognition sequence within the first exon of the human T cell receptor (TCR) alpha constant region gene. The engineered meganucleases can exhibit at least one optimized characteristic, such as enhanced (i.e., increased) specificity or efficiency of cleavage, when compared to the first-generation meganuclease TRC 1-2x.87EE. The present invention also encompasses methods of using such engineered nucleases to make genetically-modified cells, and the use of such cells in a pharmaceutical composition and in methods for treating diseases, such as cancer.

The invention includes engineered nucleases that recognize and cleave a recognition sequence within the first exon of the human T cell receptor (TCR) alpha constant region gene. The engineered meganucleases can exhibit at least one optimized property, such as enhanced (i.e., increased) specificity or cleavage efficiency, as compared to the first generation meganuclease TRC 1-2x.87EE. The invention also includes methods of using such engineered nucleases to make genetically modified cells, as well as uses of such cells in pharmaceutical compositions and in methods of treating diseases such as cancer.

The present invention provides engineered wide-range nuclease enzymes derived from I-CreI having substitutions at specific positions that increase the activity of the nuclease on recognition sequences comprising certain central sequences. The invention also provides methods of cleaving double stranded DNA using such engineered wide range nuclease enzymes. The invention further provides methods for improving the activity of engineered broad-range nuclease enzymes against recognition sequences comprising certain central sequences.

A dual vector system for treating citrullinemia is provided. The system includes (a) a gene editing AAV comprising a first AAV rh79 capsid and a first vector genome comprising a 5' ITR, a sequence encoding a meganuclease that targets PCSK9 under control of regulatory sequences that direct expression of the meganuclease in a target cell comprising a PCSK9 gene, and a 3' ITR; and (b) a donor AAV vector comprising a second AAV capsid and a second AAV rh79 vector genome comprising: a 5'ITR, a 5' homology directed recombination (HDR) arm, a transgene encoding Argininosuccinate Synthase 1 (ASS1) and regulatory sequences that direct expression of the transgene in the target cell, a 3' HDR arm, and a 3' ITR.

To provide an engineered I-CreI-derived meganuclease for recognition and cleavage of a recognition sequence having a specific center sequence.SOLUTION: One aspect is an engineered meganuclease that binds to and cleaves a recognition sequence comprising a center sequence consisting of ACAA, ACAG, ACAT, ACGA, ACGC, ACGG, ACGT, ATAA, ATAG, ATAT, ATGA, ATGG, TTGG, GCAA, GCAT, GCGA, GCAG, TCAA or TTAA, where the engineered meganuclease comprises a first subunit and a second subunit, where the first subunit and the second subunit each comprise a specific amino acid sequence with substitution at one or more positions corresponding to positions 48, 50, 71, 72, 73 and 74 of a specific amino acid sequence.SELECTED DRAWING: None

Embodiments of the present disclosure are directed to methods and compositions for treating recurrent Herpes simplex virus (HSV) disease associated with latent Herpes simplex virus type I(HSV-1) reactivation in a subject. In some embodiments, the composition comprises a plurality of at least three different adeno-associated viral vector (AAV) serotypes, each viral vector serotype comprising a first nucleic acid sequence encoding a first HSV-1-specific meganuclease, and a second nucleic acid sequence encoding a second HSV-1-specific meganuclease, separately. In some embodiments, the composition comprises an adeno-associated viral vector serotype comprising at least one sequence encoding an HSV-1-specific meganuclease targeting a duplicated region of the HSV-1 genome. In certain embodiments the adeno-associated viral vectors disclosed herein further comprise a neuron-specific promoter.

The present disclosure encompasses engineered meganucleases that bind and cleave recognition sequences within a C9Orf72 gene. The present disclosure also encompasses methods of using such engineered meganucleases to make genetically modified cells. Further, the disclosure encompasses pharmaceutical compositions comprising engineered meganuclease proteins, or polynucleotides encoding engineered meganucleases of the disclosure, and the use of such compositions for the modification of a C9Orf72 gene in a subject, or for treatment of amyotrophic lateral sclerosis (ALS) and / or frontotemporal dementia (FTD).

Disclosed are rationally-designed, non-naturally-occurring meganucleases in which a pair of enzyme subunits having specificity for different recognition sequence half-sites are joined into a single polypeptide to form a functional heterodimer with a non-palindromic recognition sequence. The invention also relates to methods of producing such meganucleases, and methods of producing recombinant nucleic acids and organisms using such meganucleases.

The present invention encompasses engineered nucleases which recognize and cleave a recognition sequence within a Hepatitis B virus (HBV) genome. The engineered meganucleases can exhibit at least one optimized characteristic, such as enhanced specificity and / or efficiency of indel formation, when compared to previously described HBV meganucleases. Further, the invention encompasses pharmaceutical compositions comprising engineered meganuclease proteins, nucleic acids encoding engineered meganucleases, and the use of such compositions for treating HBV infections or hepatocellular carcinoma.

A dual vector system for treating phenylketonuria is provided. The system includes (a) a gene editing AAV comprising a first AAV rh79 capsid and a first vector genome comprising a 5' ITR, a sequence encoding a meganuclease that targets PCSK9 under control of regulatory sequences that direct expression of the meganuclease in a target cell comprising a PCSK9 gene, and a 3' ITR; and (b) a donor AAV vector comprising a second AAV capsid and a second AAV rh79 vector genome comprising: a 5'ITR, a 5' homology directed recombination (HDR) arm, a transgene encoding phenylalanine hydroxylase (PAH) and regulatory sequences that direct expression of the transgene in the target cell, a 3' HDR arm, and a 3' ITR.

The present invention encompasses engineered nucleases which recognize and cleave a recognition sequence within a Hepatitis B virus (HBV) genome. The engineered meganucleases can exhibit at least one optimized characteristic, such as enhanced specificity and / or efficiency of indel formation, when compared to previously described HBV meganucleases. Further, the invention encompasses pharmaceutical compositions comprising engineered meganuclease proteins, nucleic acids encoding engineered meganucleases, and the use of such compositions for treating HBV infections or hepatocellular carcinoma.

The present disclosure encompasses engineered meganucleases that comprise two subunits connected by a polypeptide linker that is smaller than polypeptide linkers used in the art. The engineered meganucleases can exhibit at least one improved characteristic, such as increased efficiency of binding and / or cleavage, when compared to meganucleases comprising longer polypeptide linkers. The present disclosure also encompasses methods of using engineered nucleases to make genetically-modified cells and the use of such cells in a pharmaceutical composition and in methods for treating diseases.

Targeted transcriptional effectors (transcription activators and transcription repressors) derived from meganucleases are described. Also described are nucleic acids encoding same, and methods of using same to regulate gene expression. The targeted transcriptional effectors can comprise (i) a meganuclease DNA-binding domain lacking endonuclease cleavage activity that binds to a target recognition site; and (ii) a transcription effector domain.