Abstract: The present invention provides RNA-guided endonucleases, which are engineered for expression in eukaryotic cells or embryos, and methods of using the RNA-guided endonuclease for targeted genome modification in eukaryotic cells or embryos. Also provided are fusion proteins, wherein each fusion protein comprises a CRISPR/Cas-like protein or fragment thereof and an effector domain. The effector domain can be a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Also provided are methods for using the fusion proteins to modify a chromosomal sequence or regulate expression of a chromosomal sequence.
Abstract: Processes, oligonucleotides, and kits for amplifying RNA. In particular, the processes generate and amplify cDNA libraries in which the orientation of the input RNA molecule is preserved in the products. Among the various aspects of the present disclosure is the provision of process for directionally amplifying RNA. The process comprises reverse transcribing at least one RNA molecule in the presence of a plurality of first synthesis primers to generate a plurality of first strands of complementary DNA (cDNA), wherein each of the first synthesis primers comprises a 3? sequence having complementarity to a portion of the RNA molecule, a non-complementary 5? sequence corresponding to one or more amplification primers, and optionally an internal tag sequence comprising a first tag sequence.
Type:
Grant
Filed:
March 24, 2016
Date of Patent:
July 14, 2020
Assignee:
Sigma-Aldrich Co. LLC
Inventors:
Kenneth B Heuermann, Carol A Kreader, Jaime K Robert, Brian W Ward
Abstract: The present invention provides RNA-guided endonucleases, which are engineered for expression in eukaryotic cells or embryos, and methods of using the RNA-guided endonuclease for targeted genome modification in eukaryotic cells or embryos. Also provided are fusion proteins, wherein each fusion protein comprises a CRISPR/Cas-like protein or fragment thereof and an effector domain. The effector domain can be a cleavage domain, an epigenetic modification domain, a transcriptional activation domain, or a transcriptional repressor domain. Also provided are methods for using the fusion proteins to modify a chromosomal sequence or regulate expression of a chromosomal sequence.
Abstract: Compositions and methods for using nucleosome interacting protein domains to increase accessibility of programmable DNA modification proteins to target chromosomal sequences, thereby increasing efficiency of targeted genome/epigenetic modification in eukaryotic cells.
Type:
Grant
Filed:
July 10, 2018
Date of Patent:
March 31, 2020
Assignee:
Sigma-Aldrich Co. LLC
Inventors:
Fuqiang Chen, Xiao Ding, Yongmei Feng, Gregory D. Davis
Abstract: The present disclosure provides compositions and methods for acetylating histones at targeted chromosomal locations in a cell. In particular, the disclosure provides a fusion protein comprising a DNA binding domain and at least one histone acetyltransferase (HAT) domain, such that the DNA binding domain targets the fusion protein to a targeted chromosomal location and the HAT domain acetylates histones at the targeted location.
Abstract: Paired CRISPR nickase ribonucleoproteins engineered to target immune-related genomic loci and methods of using said ribonucleoproteins to modify the immune-related genomic loci.
Type:
Application
Filed:
April 12, 2019
Publication date:
October 17, 2019
Applicant:
SIGMA-ALDRICH CO. LLC
Inventors:
Qingzhou Ji, Gregory D. Davis, Jacob T. Lamberth
Abstract: A composition, method and device for the preparation of biological samples for subsequent instrumental analyses, such as GC, GC-MS, LC and LC-MS analysis, using a solid phase extraction (SPE) process is described. Through SPE process alone or an integrated combination of protein precipitation, filtration, and SPE using a hydrophobic zirconia-coated chromatographic media, interfering compounds, such as proteins, glycerides and phosphate-containing compounds, are eliminated from the biological, food, environmental and biotechnology samples, affording an enhanced analyte response during the instrumental analysis.
Type:
Grant
Filed:
May 21, 2010
Date of Patent:
October 8, 2019
Assignee:
Sigma-Aldrich Co. LLC
Inventors:
Xiaoning Lu, Olga I. Shimelis, Maochun M. Ye
Abstract: The present invention provides mammalian cell lines that have been genetically engineered causing such cell lines to be resistant to viral entry and/or propagation, and provides methods of using said cell lines to reduce or prevent viral contamination of biologic production systems.
Type:
Grant
Filed:
March 3, 2015
Date of Patent:
August 13, 2019
Assignee:
SIgma-Aldrich Co. LLC
Inventors:
Nan Lin, Joaquina Mascarenhas, Audrey Chang, David Onions, Henry George, Kevin Kayser
Abstract: Methods for visualizing modified nucleotides in a specific nucleic acid sequence or specific nucleic acid sequence interactions in single cells, wherein the methods comprise coupling an in situ hybridization (ISH) reaction with a proximity ligation assay (PLA) reaction.
Abstract: Methods are provided herein for identifying rare and/or unknown DNA sequences by next-generation sequencing approaches. Isolated double-stranded (ds), single-stranded (ss), or ds/ss DNA is fragmented and the fragments are polished, phosphorylated, and tailed, as necessary. Fragmentation can be enzymatic or mechanical. A universal adapter sequence is ligated to each fragment, wherein the adapter can have a top strand without a 5? phosphate, a 3? with an —H in place of the —OH, and/or a 3? extra base complementary to any base added to the polished fragments. The ligatamers may then serve as templates for ampli?cation using a forward primer complementary to the adapter sequence and a reverse primer targeted to the fragment sequence. Compositions produced by these methods and kits adapted for performing these methods are also described herein.
Abstract: Compositions and methods for using programmable DNA binding proteins to increase the efficiency and/or specificity of targeted genome modification or to facilitate the detection of specific genomic loci in eukaryotic cells.
Abstract: Methods for visualizing modified nucleotides in a specific nucleic acid sequence or specific nucleic acid sequence interactions in single cells, wherein the methods comprise coupling an in situ hybridization (ISH) reaction with a proximity ligation assay (PLA) reaction.
Abstract: The present disclosure provides reagents and methods for molecular proximity detection of specific endogenous nucleic acids in situ using RNA-guided nucleic acid binding proteins.
Type:
Application
Filed:
September 23, 2016
Publication date:
November 29, 2018
Applicant:
SIGMA-ALDRICH CO. LLC
Inventors:
Gregory D Davis, Vikas B Palhan, Carol A. Kreader
Abstract: This disclosure provides methods and devices for quantitating, separating and/or detecting water in a liquid, gas or solid sample comprising one or more chemicals, the method comprising: providing the liquid, gas or solid sample comprising water and the one or more chemicals; and exposing said liquid, gas or solid sample to at least one solid support including at least one dicationic and/or tricationic species of Formula I or II adsorbed, absorbed or immobilized on the solid support.
Abstract: Processes, oligonucleotides, and kits for amplifying RNA. In particular, the processes generate and amplify cDNA libraries in which the orientation of the input RNA molecule is preserved in the products. Among the various aspects of the present disclosure is the provision of process for directionally amplifying RNA. The process comprises reverse transcribing at least one RNA molecule in the presence of a plurality of first synthesis primers to generate a plurality of first strands of complementary DNA (cDNA), wherein each of the first synthesis primers comprises a 3? sequence having complementarity to a portion of the RNA molecule, a non-complementary 5? sequence corresponding to one or more amplification primers, and optionally an internal tag sequence comprising a first tag sequence.
Type:
Application
Filed:
March 24, 2016
Publication date:
February 15, 2018
Applicant:
Sigma-Aldrich Co. LLC
Inventors:
Kenneth B Heuermann, Carol A Kreader, Jaime K Robert, Brian W Ward