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1328 results about "Fluorescent protein" patented technology
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System and Method for Localization of Large Numbers of Fluorescent Markers in Biological Samples
ActiveUS20120193530A1Reduce respective collection solid angleExtensive collectionMaterial analysis using wave/particle radiationElectric discharge tubesFluorescenceQuantum dot
A method and system for the imaging and localization of fluorescent markers such as fluorescent proteins or quantum dots within biological samples is disclosed. The use of recombinant genetics technology to insert “reporter” genes into many species is well established. In particular, green fluorescent proteins (GFPs) and their genetically-modified variants ranging from blue to yellow, are easily spliced into many genomes at the sites of genes of interest (GoIs), where the GFPs are expressed with no apparent effect on the functioning of the proteins of interest (PoIs) coded for by the GoIs. One goal of biologists is more precise localization of PoIs within cells. The invention is a method and system for enabling more rapid and precise PoI localization using charged particle beam-induced damage to GFPs. Multiple embodiments of systems for implementing the method are presented, along with an image processing method relatively immune to high statistical noise levels.
Owner:FEI CO
Monomeric and dimeric fluorescent protein variants and methods for making same
InactiveUS7157566B2Reduced propensity to oligomerizeReduces or eliminates the degree of oligomerization of said fluorescent proteinUltrasonic/sonic/infrasonic diagnosticsFungiCrystallographyFluorescence
The present invention relates generally to variant fluorescent proteins, and more specifically to monomeric and dimeric forms of Anthozoan fluorescent proteins. In one aspect, the present invention provides variants of fluorescent proteins, where the variants have a reduced propensity to tetramerize, and form dimeric or monomeric structures. The invention also relates to methods of making and using such fluorescent protein monomers and dimers.
Owner:HOWARD HUGHES MEDICAL INST +1
T-cell epitope identification
ActiveUS20180052176A1Efficient removalMicrobiological testing/measurementBiological testingNucleotideMembrane bound
The present invention is a method for determining the identity of the epitopes recognized by T-cells. The method consists of expressing an encoded library of candidate epitope sequences in a recipient reporter cell capable of providing a detectable signal upon cytotoxic attack from a single cognate T-cell followed by contacting the reporter cells with T-cells of interest. The reporter cells with a single indicating cytotoxic attack from a T-cell are isolated and then analyzed by next-generation sequencing in order to identify the epitope sequences. Specifically disclosed is a method in which a library of candidate epitope-encoding nucleic acids are expressed in cells which feature a membrane-bound major histocompatibility complex (MHC) protein, said library produced by transfection of plasmids featuring both a nucleotide encoding the candidate epitope and a nucleotide encoding a FRET-based fluorescent protein cleaved by granzyme.
Owner:PROVINCIAL HEALTH SERVICES AUTHORITY
Methods and compositions comprising Renilla GFP
InactiveUS7528242B2Antibody mimetics/scaffoldsFusion with protease sitePtilosarcusFluorescent protein
Owner:RIGEL PHARMA INC
Method and computer program product for distinguishing and sorting seeds containing a genetic element of interest
InactiveUS20090032441A1Speed up the processMicrobiological testing/measurementInvestigating moving fluids/granular solidsBulk samplesFluorescent protein
Owner:PIONEER HI BRED INT INC
CAR-T transgene vector based on replication defective recombinant lentivirus and construction method and application of CAR-T transgene vector
ActiveCN105602992ASignificant effectPromote secretionGenetic material ingredientsFermentationEucaryotic cellAmpicillin
The invention discloses a CAR-T transgene vector based on replication defective recombinant lentivirus. The CAR-T transgene vector comprises an original nuclear replicon pUCOri sequence, a resistance gene AmpR sequence containing ampicillin, a virus replicon SV40 Ori sequence, a lentivirus packaging cis element, ZsGreen1 green fluorescent protein, an IRES ribosome binding sequence, a human EF1 alpha promoter , a chimeric antigen receptor of second-generation CAR or third-generation CAR and a regulating element, wherein the original nuclear replicon pUCOri sequence is used for plasmid replication; the resistance gene AmpR sequence is used for massively proliferating target strains; the virus replicon SV40 Ori sequence is used for enhancing replication in eukaryocyte; the lentivirus packaging cis element is used for lentivirus packaging; the ZsGreen1 green fluorescent protein is used for expressing green fluorescent for eukaryocyte; the IRES ribosome binding sequence is used for jointly transcribing and expressing protein; the human EF1 alpha promoter is used for conducting eukaryotic transcription on antigen receptor genes; the chimeric antigen receptor is used for forming the second-generation CAR or the third-generation CAR integrating recognition, transfer and start; the regulating element is used for enhancing expression efficiency of transgenes and used after eWPRE-enhanced type woodchuck hepatitis b virus is transcribed. Besides, the invention further discloses a construction method and application of the vector. By means of the CAR-T transgene vector and the construction method and application of the vector, secretion of cell factors and an in vitro killing effect of CAR-T cells can be remarkably improved, and the clinical treatment effect is remarkable.
Owner:SHANGHAI UNICAR THERAPY BIOPHARM TECH CO LTD
Oxidation reduction sensitive green fluorescent protein variants
The disclosure provides proteins that can be used to determine the redox status of an environment (such as the environment within a cell or subcellular compartment). These proteins are green fluorescent protein (GFP) variants (also referred to as redox sensitive GFP (rosGFP) mutants), which have been engineered to have two cysteine amino acids near the chromophore and within disulfide bonding distance of each other. Also provided are nucleic acid molecules that encode rosGFPs, vectors containing such encoding molecules, and cells transformed therewith. The disclosure further provides methods of using the rosGFPs (and encoding molecules) to analyze the redox status of an environment, such as a cell, or a subcellular compartment within a cell. In certain embodiments, both redox status and pH are analyzed concurrently.
Owner:OREGON HEALTH & SCI UNIV
Expression vector, methods for the production of heterologous gene products and for the selection of recombinant cells producing high levels of such products
ActiveUS20040148647A1Shorten the timeReduce development costsAnimal cellsMicrobiological testing/measurementHeterologousHamster
An expression vector for eukaryotic cells comprising a gene which codes for a protein of interest, functionally linked to a hamster-ubiquitin / S27a-promoter and a gene which codes for a fluorescent protein. Preferably the expression vector also contains an amplifiable selectable marker gene. The invention also describes host cells, preferably mammalian cells, which have been transfected with the expression vector, processes for producing heterologous gene products and a method of selecting high-producing cells.
Owner:BOEHRINGER INGELHEIM PHARM KG
GFP-SNAP25 fluorescence release assay for botulinum neurotoxin protease activity
The present invention provides a nucleic acid molecule which contains a nucleotide sequence encoding a SNAP-25 substrate which includes (i) a green fluorescent protein; (ii) a first partner of an affinity couple; and (iii) a portion of SNAP-25 that includes a BoNT / A, BoNT / C1 or BoNT / E recognition sequence containing a cleavage site, where the cleavage site intervenes between the green fluorescent protein and the first partner of the affinity couple. Further provided herein is a nucleic acid molecule which contains a nucleotide sequence encoding a tagged toxin substrate which includes (i) a fluorescent protein; (ii) a first partner of an affinity couple; and (iii) a clostridial toxin recognition sequence containing a cleavage site, where the cleavage site intervenes between the fluorescent protein and the first partner of the affinity couple.
Owner:ALLERGAN INC
Emission ratiometric indicators of phosphorylation by C-kinase
InactiveUS20050026234A1Facilitate its translocationPeptide/protein ingredientsAntibody mimetics/scaffoldsNucleotideFluorophore
A chimeric phosphorylation indicator (CPI) as provided herein can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. Where the phosphorylatable domain is phosphorylatable by protein kinase C (PKC), the CPI is a c-kinase activity reporter (CKAR). Donor and acceptor molecules may be, independently, fluorescent proteins such as non-oligomerizing fluorescent proteins. A CPI can contain a phosphorylatable polypeptide and a fluorescent protein; the phosphorylatable polypeptide may be contained within the sequence of the fluorescent protein, or the fluorescent protein may be contained within the sequence of the phosphorylatable polypeptide. The spatiotemporal properties of the PKC signal pathway may be tested with CKAR, calcium-sensing fluorophores and FRET-based translocation assays. Polynucleotides encoding such CPIs, and kits containing the indicators and / or the polynucleotides, are provided. A method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.
Owner:RGT UNIV OF CALIFORNIA +1
Nucleic acids encoding non aggregating fluorescent proteins and methods for using the same
Nucleic acid compositions encoding non-aggregating chromo / fluoroproteins and mutants thereof, as well as the encoded proteins, are provided. The proteins of interest are polypeptides that are non-aggregating colored and / or fluorescent proteins, where the non-aggregating feature arises from the modulation of residues in the N-terminus of the protein and the chromo and / or fluorescent feature arises from the interaction of two or more residues of the protein. Also provided are fragments of the subject nucleic acids and the peptides encoded thereby, as well as antibodies to the subject proteins and transgenic cells and organisms. The subject protein and nucleic acid compositions find use in a variety of different applications. Finally, kits for use in such applications, e.g., that include the subject nucleic acid compositions, are provided.
Owner:TAKARA BIO USA INC
Tandem fluorescent protein constructs
This invention provides tandem fluorescent protein construct including a donor fluorescent protein moiety, an acceptor fluorescent protein moiety and a linker moiety that couples the donor and acceptor moieties. The donor and acceptor moieties exhibit fluorescence resonance energy transfer which is eliminated upon cleavage. The constructs are useful in enzymatic assays.
Owner:VERTEX PHARMA SAN DIEGO LLC +1
Compositions and methods for measuring analyte concentrations
The current invention relates to fusion proteins comprising at least one functional periplasmic binding protein, at least one labeling moiety and at least one fluorescent protein. In one embodiment, the periplasmic binding protein is a functional glucose-galactose binding protein (GGBP). The invention also relates to methods for quantifying an analyte, for example glucose, in a cell or tissue comprising administering a composition comprising a fluorescent periplasmic binding fusion protein portion to the cell or tissue, and measuring the fluorescence of the fluorescent periplasmic binding fusion protein.
Owner:BECTON DICKINSON & CO
Non-linear structure light illumination microscopic imaging method and system
ActiveCN104515759AImprove imaging resolutionReduce bleaching effectFluorescence/phosphorescenceRapid imagingFluorescence
The invention discloses a non-linear structure light illumination microscopic imaging method which comprises the following steps: 1) loading a computed hologram on a digital microscopic array; 2) generating a first spatial structure light field which meets sine distribution and is used for activating fluorescent protein, and radiating the first spatial structure light field to the surface of a sample, so as to convert a part of protein to be in an illuminated state from a dark state; 3) radiating the sample in a second spatial structure light field so as to enable fluorescent protein in the bright state to emit fluorescent light, collecting the fluorescent light, and imaging in a photoelectric detector; 4) repeating the step 2) and the step 3), acquiring a plurality of spatial frequencies, acquiring a plurality of initial phases in each direction to obtain a plurality of original images, and reestablishing a super-resolution image according to a GPU acceleration algorithm. The invention further discloses a non-linear structure light illumination microscopic imaging system. The non-linear structure light illumination microscopic imaging method has the advantages of relatively high system imaging resolution, high fluorescent drifting resistance, low phototoxicity and rapid imaging.
Owner:SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
Measurement of fluorescence using capillary isoelectric focusing
Improved apparatus is disclosed for carrying out axially illuminated laser induced fluorescence whole-column imaging detection in the capillary isoelectric focusing of proteins. The separation capillary was made of low refractive index Teflon conditioned with methylcellulose to reduce electroosmotic flow and a small amount of high refractive index organic solvent (glycerol) was added to the sample mixture. It was found that an axially directed laser excitation beam was propagated essentially with total internal reflection, so that minimum interference arose from stray light or from scattering light originating from the wall of the capillary. With the naturally fluorescent protein R-phycoerythrin, a concentration detection limit LOD 10−11 M or mass LOD 10−17 Mo was obtained.
Owner:PAWLISZYN JANUSZ
Method and system for tomographic imaging using fluorescent proteins
A system for optical tomography includes an apparent light source adapted to project excitation light toward a specimen having fluorescent proteins therein, wherein the excitation light enters the specimen becoming intrinsic light within the specimen, wherein the intrinsic light is adapted to excite fluorescent light from the fluorescent proteins, and wherein the intrinsic light and the fluorescent light are diffuse. A method of optical tomography includes generating the excitation light with the apparent light source, wherein the intrinsic light and the fluorescent light are diffuse.
Owner:THE GENERAL HOSPITAL CORP
Fluorescent protein sensors for detection of analytes
InactiveUS7060869B2Reduce sensitivityPeptide/protein ingredientsAntibody mimetics/scaffoldsEnergy transferAnalyte
Owner:RGT UNIV OF CALIFORNIA
Method for targeted knockout of gene ALK6 by using CRISPR-Cas9
InactiveCN105925608AImprove universalityReduced editing efficiencyNucleic acid vectorReceptors for growth factors/regulatorsDouble strandedTransfection
The invention discloses a method for targeted knockout of a gene ALK6 by using CRISPR-Cas9. The method comprises the steps of carrying out homologous comparison on a plurality of species so as to select two pairs of gene ALK6 target sequences, synthesizing DNA double strands, of which two pairs of target sequences are different and expressed proteins are the same and which have the same BsmBI cohesive ends, by using PAM design principles of gRNA, connecting the double strands with plasmid pPDNA330 so as to obtain recombinant vectors carrying ALK6 homologous genes of a plurality of species, detecting gene knockout efficiency of the recombinant vectors by using a fluorescin expression method, carrying out transfection on a vector with higher knockout efficiency with recipient cells of a plurality of species, and carrying out gene knockout and verification, thereby completing simple, efficient and accurate gene knockout of genes ALK6 of a plurality of species.
Owner:GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
Cell line gene knock-out method for obtaining large fragment deletion through CRISPR/Cas9 system
ActiveCN107435051AEasy accessImprove knockout productivityVectorsStable introduction of DNAFluorescenceLarge fragment
The invention relates to a cell line gene knock-out method for obtaining large fragment deletion through a CRISPR / Cas9 system, and belongs to the field of genetic engineering and genetic modification. A pX458 vector is modified, so that the pX458 vector is provided with DsRed2 and ECFP (Enhanced Cyan Fluorescence Protein); then, a plurality of specific sgRNA sites are designed by aiming at a target gene and are connected into the modified vector; after the cell line transfection, cell groups are sorted by a flow cytometry; single cells subjected to genome editing can be very fast obtained; then, a single-cell DNA (Deoxyribonucleic Acid) sequence is subjected to PCR (Polymerase Chain Reaction) amplification; and single cells with large fragment deletion can be picked out from the single cells through gel electrophoresis. The CRISPR / Cas9 system, the flow cytometry single-cell sorting and fluorescent protein screening on the expression vector are combined, so that the positive monoclonal cells with the large fragment deletion can be obtained in a short time; and the work efficiency of the cell line gene knock-out is greatly improved.
Owner:XINXIANG MEDICAL UNIV
Recombinant constructs and transgenic fluorescent ornamental fish therefrom
ActiveUS20090025645A1Low densityReduce the numberVectorsData processing applicationsTransgenesisCoral
The present invention relates to the method and use of reef coral fluorescent proteins in making transgenic red, green and yellow fluorescent zebrafish. Preferably, such fluorescent zebrafish are fertile and used to establish a population of transgenic zebrafish and to provide to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed.
Owner:GLOFISH LLC
Method for knocking out MSTN (myostatin) genes in targeted manner by utilizing CRISPR-Cas9
InactiveCN106119283AImprove universalityReduced editing efficiencyNucleic acid vectorReceptors for growth factors/regulatorsMyostatinDouble strand
The invention discloses a method for knocking out MSTN (myostatin) genes in a targeted manner by utilizing CRISPR-Cas9. According to the method, multiple species are subjected to homology comparison, two pairs of MSTN gene target sequences are selected, two pairs of DNA double strands which have the sequences different from the target sequences, the same expression protein and the same BsmBi sticky ends are synthesized according to the design principle of PAM of gRNA, the double strands are connected with pPDNA330 plasmids, a recombinant vector carrying MSTN homologous genes of multiple species is obtained and is used for detecting the gene knockout efficiency with a fluorescent protein expression method, carrier-transfected receptor cells, having higher knockout efficiency, of multiple species are selected, gene knockout and verification are performed, and simple, efficient and accurate gene knockout on the MSTN genes of the multiple species is completed.
Owner:GUANGXI ZHUANG AUTONOMOUS REGION BUFFALO INST
Emission ratiometric indicators of phosphorylation
InactiveUS6900304B2Facilitate its translocationUltrasonic/sonic/infrasonic diagnosticsFungiPhosphoamino AcidsNucleotide
A chimeric phosphorylation indicator is provided. A chimeric phosphorylation indicator can contain a donor molecule, a phosphorylatable domain, a phosphoaminoacid binding domain (PAABD), and an acceptor molecule. A chimeric phosphorylation indicator also can contain a phosphorylatable polypeptide and a fluorescent protein, wherein the phosphorylatable polypeptide is contained within the sequence of the fluorescent protein, or wherein the fluorescent protein is contained within the sequence of the phosphorylatable polypeptide. Also provided are polynucleotides encoding such chimeric phosphorylation indicators, as well as kits containing the indicators or the polynucleotides. In addition, a method of using the chimeric phosphorylation indicators to detect a kinase or phosphatase in a sample is provided.
Owner:RGT UNIV OF CALIFORNIA
Vectors for plant transformation and methods of use
InactiveUS20060041956A1Easy to eliminateImprove efficiencyOther foreign material introduction processesFermentationNormal appearanceRead through
The present invention is directed to a vector for identifying read-through of non-T-DNA in a T-DNA vector. In one embodiment, the vector provides a visually detectable change in the normal appearance of transformants wherein read-through has occurred. In another embodiment, the vector also provides for expression of a readily detectable fluorescent protein that allows for the early detection and elimination of transformants wherein read-through has occurred. In a further aspect, the present invention is directed to a method for detecting read-through of non-T-DNA in plants transformed with a T-DNA vector. In another aspect, the present invention is directed to a method for producing a transgenic plant containing a polynucleotide of interest but being substantially free of non-T-DNA.
Owner:PIONEER HI BRED INT INC
Monomeric and dimeric fluorescent protein variants and methods for making same
InactiveUS20050196768A1Improve efficiencyReduction tendencyCompound screeningApoptosis detectionDimerBiochemistry
The present invention relates generally to fluorescent proteins and fluorescent protein variants, and more specifically to monomeric and dimeric forms of Anthozoan fluorescent proteins. In one aspect, the present invention provides variants of fluorescent proteins, where the variants have a reduced propensity to tetramerize, and form dimeric or monomeric structures. In a further aspect, the present invention provides variants of fluorescent proteins, the variants being characterized by more efficient maturation than corresponding fluorescent proteins from which they are derived. The invention also relates to methods of making and using such fluorescent proteins and fluorescent protein variants, including fluorescent protein monomers and dimers.
Owner:RGT UNIV OF CALIFORNIA
Method of screening multiply transformed cells using bicistronic expression of fluorescent proteins
A method of screening multiply transformed / transfected cells to identify those cells expressing at least two peptides or proteins of interest. The method comprising: 1. Simultaneously or sequentially transforming a cell with at least two different expression cassettes in which the gene of interest is linked via an IRES to a fluorescent marker gene. Each marker gene is different. 2. Providing conditions in which expression of the genes will occur. 3. Identifying cells expressing proteins by detecting the different fluorescent signals.
Owner:NEWSOUTH INNOVATIONS PTY LTD
Method for rapidly obtaining CRISPR/Cas9 (clustered regularly interspersed short palindromic repeats/Cas9) gene knockout stable cell line through monoclonal cell sorting
InactiveCN107418974AImprove knockout productivitySave the trouble of screeningVectorsStable introduction of DNAFluorescenceHigh survival rate
The invention relates to a method for rapidly obtaining a CRISPR / Cas9 (clustered regularly interspersed short palindromic repeats / Cas9) gene knockout stable cell line through monoclonal cell sorting and belongs to the technical field of genetic engineering and genetic modification. According to the method, a CRISPR / Cas9 system, single-cell sorting by a flow cytometer and fluorescent protein screening on an expression vector are combined, positive monoclonal cells can be obtained in short time, and gene knockout work efficiency of the cell line is greatly improved. Compared with a traditional cell line gene knockout method, the flow cytometer is utilized for single-cell sorting, on one hand, tedious work of antibiotic screening is omitted, so that a large quantity of single cells can be obtained in quite short time, and on the other hand, the condition that cells in each culture hole are single cells can be guaranteed and false positive rate is reduced. Sorting is performed 40-80 h after cell transfection, at the time point, the highest survival rate of the cells after sorting can be guaranteed, and the screening efficiency is improved accordingly.
Owner:XINXIANG MEDICAL UNIV
Fluorescent protein sensors for detection of analytes
InactiveUS20060112440A1Reduce sensitivitySugar derivativesAntibody mimetics/scaffoldsEnergy transferAnalyte
Fluorescent indicators including a binding protein moiety, a donor fluorescent protein moiety, and an acceptor fluorescent protein moiety are described. The binding protein moiety has an analyte-binding region which binds an analyte and causes the indicator to change conformation upon exposure to the analyte. The donor moiety and the acceptor moiety change position relative to each other when the analyte binds to the analyte-binding region. The donor moiety and the acceptor moiety exhibit fluorescence resonance energy transfer when the donor moiety is excited and the distance between the donor moiety and the acceptor moiety is small. The indicators can be used to measure analyte concentrations in samples, such as calcium ion concentrations in cells.
Owner:RGT UNIV OF CALIFORNIA
Circularly permuted fluorescent protein indicators
Polynucleotides encoding fluorescent indicators, which contain a sensor polypeptide inserted within a fluorescent moiety, are provided, as are polypeptides encoded by such polynucleotides. Also provided are circularly permuted fluorescent polypeptides and polynucleotides encoding the circularly permuted fluorescent polypeptides. In addition, methods of using the fluorescent indicators and the circularly permuted fluorescent polypeptides are provided.
Owner:HOWARD HUGHES MEDICAL INST +1
Renilla reniformis fluorescent proteins, nucleic acids encoding the fluorescent proteins and the use thereof in diagnostics, high throughput screening and novelty items
Isolated and purified nucleic acids encoding green fluorescent proteins from Renilla reniformis and the green fluorescent protein encoded thereby are also provided. Mutants of the nucleic acid molecules and the modified encoded proteins are also provided. Compositions and combinations comprising the green fluorescent proteins and / or the luciferase are further provided.
Owner:PROLUME +2
Fluorescent protein variants and methods for making same
InactiveUS7005511B2Reduced propensity to oligomerizeReduces or eliminates the degree of oligomerization of said fluorescent proteinFungiBacteriaCrystallographyFluorescence
The present invention relates generally to variant fluorescent proteins, and more specifically to monomeric and dimeric forms of Anthozoan fluorescent proteins. In one aspect, the present invention provides variants of fluorescent proteins, where the variants have a reduced propensity to tetramerize, and form dimeric or monomeric structures. The invention also relates to methods of making and using such fluorescent protein monomers and dimers.
Owner:HOWARD HUGHES MEDICAL INST +1
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