Abstract: The invention provides compositions and methods for selectively reducing the expression of a gene product from a desired target gene, as well as treating diseases caused by expression of the gene. The method involves introducing into the environment of a cell an amount of a double-stranded RNA (dsRNA) such that a sufficient portion of the dsRNA can enter the cytoplasm of the cell to cause a reduction in the expression of the target gene. The dsRNA has a first oligonucleotide sequence that is between 26 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of from about 19 to about 23 nucleotides is complementary to a nucleotide sequence of the RNA produced from the target gene.
Type:
Application
Filed:
June 13, 2016
Publication date:
November 24, 2016
Applicants:
CITY OF HOPE, INTEGRATED DNA TECHNOLOGIES, INC.
Inventors:
John J. ROSSI, Mark A. BEHLKE, Dongho KIM
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Grant
Filed:
November 10, 2014
Date of Patent:
September 13, 2016
Assignees:
CITY OF HOPE, INTEGRATED DNA TECHNOLOGIES, INC.
Inventors:
John J. Rossi, Mark A. Behlke, Dongho Kim
Abstract: This invention pertains to modified compositions for use in CRISPR systems, and their methods of use. In particular, length-modified and chemically-modified forms of crRNA and tracrRNA are described for use as a reconstituted guide RNA for interaction with Cas9 of CRIPSR systems. The resultant length-modified and chemically-modified forms of crRNA and tracrRNA are economical to produce and can be tailored to have unique properties relevant to their biochemical and biological activity in the context of the CRIPSR Cas9 endonuclease system.
Type:
Application
Filed:
December 18, 2015
Publication date:
June 23, 2016
Applicant:
INTEGRATED DNA TECHNOLOGIES, INC.
Inventors:
Michael Allen Collingwood, Ashley Mae Jacobi, Garrett Richard Rettig, Mollie Sue Schubert, Mark Aaron Behlke
Abstract: The invention provides compositions and methods for selectively reducing the expression of a gene product from a desired target gene, as well as treating diseases caused by expression of the gene. The method involves introducing into the environment of a cell an amount of a double-stranded RNA (dsRNA) such that a sufficient portion of the dsRNA can enter the cytoplasm of the cell to cause a reduction in the expression of the target gene. The dsRNA has a first oligonucleotide sequence that is between 26 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of from about 19 to about 23 nucleotides is complementary to a nucleotide sequence of the RNA produced from the target gene.
Type:
Grant
Filed:
September 6, 2013
Date of Patent:
June 14, 2016
Assignees:
Integrated DNA Technologies, Inc., City of Hope
Inventors:
John J. Rossi, Mark A. Behlke, Dongho Kim
Abstract: This invention relates to primers and probes for detecting Ebola virus and one or more subtypes of Ebola virus as well as kits including the probes and primers and methods of using the probes and primers.
Abstract: A composition comprising an oligonucleotide having the structure 5?-Y1-L1-X-L2-Y2-3?. Y1 comprises a sequence of one or more DNA or RNA nucleotides, including a first nucleotide N1 having a 3? phosphate covalently linked to L1. Y2 comprises a sequence of one or more DNA or RNA nucleotides, including a second nucleotide N2 having a 5? phosphate covalently linked to L2. L1 and L2 each independently are a direct bond or a C1-C7 alkyl, alkynyl, alkenyl, heteroalkyl, substituted alkyl, aryl, heteroaryl, substituted aryl, cycloalkyl, alkylaryl, or alkoxyl group. X is R1 is a hydrogen or a C1-C8 alkyl. M is a label or ligand comprising a fused polycyclic aromatic moiety.
Type:
Grant
Filed:
December 20, 2014
Date of Patent:
January 12, 2016
Assignee:
Integrated DNA Technologies, Inc.
Inventors:
Scott Rose, Mark A. Behlke, Richard Owczarzy, Joseph A. Walder, Derek M. Thomas, Michael R. Marvin
Abstract: The present invention provides methods that more accurately predict melting temperatures for duplex oligomers. The invented methods predict the Tm of chimeric duplexes containing various amounts of locked nucleic acid modifications in oligonucleotide strands.
Type:
Grant
Filed:
July 29, 2011
Date of Patent:
July 14, 2015
Assignee:
Integrated DNA Technologies, Inc.
Inventors:
Mark Behlke, Richard Owczarzy, Scott D. Rose, Andrey Tataurov, Yong You
Abstract: This invention pertains to improved methods for the synthesis of long, double stranded nucleic acid sequences containing difficult to clone or variable regions.
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Application
Filed:
November 10, 2014
Publication date:
April 23, 2015
Applicants:
CITY OF HOPE, INTEGRATED DNA TECHNOLOGIES, INC.
Inventors:
John J. ROSSI, Mark A. BEHLKE, Dongho KIM
Abstract: A composition comprising an oligonucleotide having the structure 5?-Y1-L1-X-L2-Y2-3?. Y1 comprises a sequence of one or more DNA or RNA nucleotides, including a first nucleotide N1 having a 3? phosphate covalently linked to L1. Y2 comprises a sequence of one or more DNA or RNA nucleotides, including a second nucleotide N2 having a 5? phosphate covalently linked to L2. L1 and L2 each independently are a direct bond or a C1-C7 alkyl, alkynyl, alkenyl, heteroalkyl, substituted alkyl, aryl, heteroaryl, substituted aryl, cycloalkyl, alkylaryl, or alkoxyl group. X is R1 is a hydrogen or a C1-C8 alkyl. M is a label or ligand comprising a fused polycyclic aromatic moiety.
Type:
Grant
Filed:
March 28, 2011
Date of Patent:
December 23, 2014
Assignee:
Integrated DNA Technologies, Inc.
Inventors:
Scott Rose, Mark A. Behlke, Richard Owczarzy, Joseph A. Walder, Derek M. Thomas, Michael R. Marvin
Abstract: The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3? end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.
Type:
Grant
Filed:
July 22, 2009
Date of Patent:
December 16, 2014
Assignee:
Integrated DNA Technologies, Inc.
Inventors:
Joseph Alan Walder, Mark Aaron Behlke, Scott Rose, Joseph Dobosy
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Application
Filed:
August 19, 2014
Publication date:
December 4, 2014
Applicants:
CITY OF HOPE, INTEGRATED DNA TECHNOLOGIES, INC.
Inventors:
John J. ROSSI, Mark A. BEHLKE, Dongho KIM
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Grant
Filed:
June 8, 2012
Date of Patent:
November 11, 2014
Assignees:
City of Hope, Integrated DNA Technologies, Inc.
Inventors:
John J. Rossi, Mark A. Behlke, Dongho Kim
Abstract: The invention provides a provides improvements to assays that employ RNase H cleavage for biological applications related to nucleic acid amplification and detection, where the RNase H has been reversibly inactivated.
Type:
Application
Filed:
March 15, 2013
Publication date:
September 11, 2014
Applicant:
Integrated DNA Technologies
Inventors:
Joseph Alan WALDER, Mark Aaron BEHLKE, Scott D. ROSE, Joseph DOBOSY, Susan Marie RUPP
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Grant
Filed:
July 8, 2011
Date of Patent:
August 19, 2014
Assignees:
City of Hope, Integrated DNA Technologies, Inc.
Inventors:
John J. Rossi, Mark A. Behlke, Dongho Kim
Abstract: The invention is directed to compositions and methods for selectively reducing the expression of a gene product from a desired target gene in a cell, as well as for treating diseases caused by the expression of the gene. More particularly, the invention is directed to compositions that contain double stranded RNA (“dsRNA”), and methods for preparing them, that are capable of reducing the expression of target genes in eukaryotic cells. The dsRNA has a first oligonucleotide sequence that is between 25 and about 30 nucleotides in length and a second oligonucleotide sequence that anneals to the first sequence under biological conditions. In addition, a region of one of the sequences of the dsRNA having a sequence length of at least 19 nucleotides is sufficiently complementary to a nucleotide sequence of the RNA produced from the target gene to trigger the destruction of the target RNA by the RNAi machinery.
Type:
Grant
Filed:
July 6, 2011
Date of Patent:
August 5, 2014
Assignees:
City of Hope, Integrated DNA Technologies, Inc.
Inventors:
John J. Rossi, Mark A. Behlke, Dongho Kim
Abstract: The present invention relates to a rapid detection of microbial-associated nuclease activity with chemically modified nuclease (e.g., ribonuclease) substrates, and probes and compositions useful in detection assays. Accordingly, in certain embodiments, the present invention provides a probe for detecting a microbial endonuclease comprising a substrate oligonucleotide of 2-30 nucleotides in length, a fluorescence-reporter group operably linked to the oligonucleotide, and a fluorescence-quencher group operably linked to the oligonucleotide. The fluorescence-reporter group and the fluorescence-quencher group are separated by at least one RNAse-cleavable residue, e.g., RNA base.
Type:
Application
Filed:
August 30, 2012
Publication date:
July 17, 2014
Applicants:
INTEGRATED DNA TECHNOLOGIES, INC., UNIVERSITY OF IOWA RESEARCH FOUNDATION
Inventors:
James O. McNamara, II, Katie R. Flenker, Lingyan Huang, Alexander R. Horswill, Mark A. Behlke, Frank J. Hernandez
Abstract: The present invention pertains to novel oligonucleotide compounds for use in various biological assays, such as nucleic acid amplification, ligation and sequencing reactions. The novel oligonucleotides comprise a ribonucleic acid domain and a blocking group at or near the 3? end of the oligonucleotide. These compounds offer an added level of specificity previously unseen. Methods for performing nucleic acid amplification, ligation and sequencing are also provided. Additionally, kits containing the oligonucleotides are also disclosed herein.
Type:
Application
Filed:
August 5, 2013
Publication date:
April 17, 2014
Applicant:
INTEGRATED DNA TECHNOLOGIES
Inventors:
Joseph Alan Walder, Mark Aaron Behlke, Scott Rose, Joesph Dobosy