Abstract: The present invention relates to modification of RNA with 5?-cap analogs in order to improve the stability and increase the expression of said RNA, in particular in immature antigen presenting cells. The present invention provides a vaccine composition comprising said stabilized RNA, immature antigen presenting cells comprising said stabilized RNA, and methods for stimulating and/or activating immune effector cells and for inducing an immune response in an individual using said stabilized RNA.
Type:
Application
Filed:
August 3, 2010
Publication date:
August 2, 2012
Applicant:
OCV Intellectual Capital , LLC
Inventors:
Ugur Sahin, Andreas Kuhn, Edward Darzynkiewicz, Jacek Jemielity, Joanna Kowalska
Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
Type:
Application
Filed:
May 11, 2005
Publication date:
September 18, 2008
Inventors:
Anthony P. Albino, Ellen D. Jorgensen, Frank Traganos, Zbigniew Darzynkiewicz, Wendy Jin
Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
Type:
Application
Filed:
February 5, 2010
Publication date:
June 3, 2010
Applicant:
VECTOR TOBACCO, INC.
Inventors:
Anthony P. Albino, Ellen D. Jorgensen, Frank Traganos, Zbigniew Darzynkiewicz, Wendy Jin
Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Application
Filed:
September 29, 2004
Publication date:
February 24, 2005
Inventors:
Zbigniew Darzynkiewicz, Frank Traganos, Gloria Juan, Stefan Gruenwald
Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
Type:
Application
Filed:
March 28, 2011
Publication date:
September 8, 2011
Applicants:
VECTOR TOBACCO, INC., New York Medical College
Inventors:
Anthony P. Albino, Ellen D. Jorgensen, Frank Traganos, Zbigniew Darzynkiewicz, Wendy Jin
Abstract: The primary and/or secondary structure of nucleic acid as well as its content and molecular weight can be analyzed in solutions by light scatter or fluorescence measurements after treatment with 3 or 4 ring aromatic cations which bind to single-stranded nucleic acids by cooperative association and induce their condensation (collapse). Preparative separation of nucleic acids of different types from the mixtures in the solution is also accomplished by the same principle and techniques wherein each condensed acid is removed from solution and the condensation reversed.
Type:
Grant
Filed:
May 23, 1984
Date of Patent:
January 31, 1989
Assignee:
Sloan-Kettering Institute for Cancer Research
Inventors:
Jan Kapuscinski, Zbigniew Darzynkiewicz
Abstract: Aspects of the invention concern methods for detecting, identifying and evaluating tobacco and tobacco products to determine the potential that these compositions have to contribute to a tobacco-related disease. It is based, at least in part; on the discovery that exposure of pulmonary cells to smoke or smoke condensate obtained from tobacco or tobacco products induces double stranded breaks in cellular DNA, which were efficiently detected using assays that measure the presence, absence, or amount of phosphorylation of the histone, H2AX.
Type:
Grant
Filed:
May 11, 2005
Date of Patent:
February 16, 2010
Assignee:
Vector Tobacco, Inc.
Inventors:
Anthony P. Albino, Ellen D. Jorgensen, Frank Traganos, Zbigniew Darzynkiewicz, Wendy Jin
Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Application
Filed:
February 24, 1999
Publication date:
June 12, 2003
Inventors:
ZBIGNIEW DARZYNKIEWICZ, FRANK TRAGANOS, GLORIA JUAN, STEFAN GRUENWALD
Abstract: Dinucleotide cap analogs are disclosed, modified at different phosphate positions with a boranophosphate group or a phosphoroselenoate group. The analogs are useful as reagents in the preparation of capped mRNAs and have increased stability both in vitro and in vivo. They may be used as inhibitors of cap-dependent translation. Optionally, the boranophosphate or phosphoroselenoate group has a 2?-O or 3?-O-alkyl group, preferably a methyl group, producing analogs called BH3-ARCAs or Se-ARCAs. ARCAs may be modified with ?-, ?-, or ?-boranophosphate or phosphoroselenoate groups.
Type:
Grant
Filed:
June 4, 2009
Date of Patent:
August 27, 2013
Assignee:
Board of Supervisors of Louisiana State University And Agricultural and Mechanical College
Inventors:
Joanna Kowalska, Jacek Jemielity, Edward Darzynkiewicz, Robert E. Rhoads, Maciej Lukaszewicz, Joanna Zuberek
Abstract: The invention provides novel affinity labels for Ser proteases of formula: L-A-X—NH—CH(R?)C(?O)CH2Cl??(I) wherein L, A, X, and R? have any of the values defined in the specification, or salts thereof, as well as compositions comprising such compounds or salts. The composition of the amino acid side-chain (R?) along with the amino acid or amino acid sequence (peptide) of the X component of formula I, affect the target selectivity of the labeled affinity ligand. Utilization of cell permeable, enzyme selective, labeled affinity ligands, provides a precise mechanism for evaluating the current and future status of cell populations.
Type:
Application
Filed:
June 21, 2004
Publication date:
November 22, 2007
Inventors:
David Phelps, Gary Johnson, Brian Lee, Zbigniew Darzynkiewicz, Jerzy Grabarek
Abstract: A process for characterizing sperm motility and viability by staining a sperm sample with Rhodamine 123 and ethidium bromide, and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm, the green counts being correlated with sperm motility and the red counts being correlated with putative dying or dead cells. Additionally, a sample of sperm can be characterized as to type and normality by staining a sample of sperm with acridine orange and simultaneously measuring the sperm fluorescence emissions at green frequencies 515-575 nm and at red frequencies 600-650 nm.
Type:
Grant
Filed:
September 1, 1982
Date of Patent:
December 17, 1985
Assignee:
Memorial Sloan Kettering Cancer Center
Inventors:
Donald P. Evenson, Zbigniew Darzynkiewicz
Abstract: The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as Serine proteases, can be key determinates in assessing; 1) the presence of tumor (cancer) cells, 2) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, and 5) the disease state status of a cell. The identification of the up or down regulation relationships of serine proteases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future physiological state of these cell populations.
Type:
Application
Filed:
June 21, 2004
Publication date:
July 6, 2006
Inventors:
David Phelps, Gary Johnson, Brian Lee, Zbigniew Darzynkiewicz, Jerzy Grabarek
Abstract: The present invention relates to modification of RNA with 5?-cap analogs of Formula (1): wherein R1-R6 and n are as described herein, in order to improve the stability and increase the expression of said RNA, in particular in immature antigen presenting cells. The present invention provides a vaccine composition comprising said stabilized RNA, immature antigen presenting cells comprising said stabilized RNA, and methods for stimulating and/or activating immune effector cells and for inducing an immune response in an individual using said stabilized RNA.
Type:
Grant
Filed:
August 3, 2010
Date of Patent:
March 29, 2016
Assignees:
BIONTECH AG, TRON-TRANSLATIONALE ONKOLOGIE AN DER UNIVERSITATSMEDIZIN DER JOHANNES GUTENBERG-UNIVERSITAT MAINZ GEMEINNUTZIGE GMBH, UNIWERSYTET WARSZAWSKI
Inventors:
Ugur Sahin, Andreas Kuhn, Edward Darzynkiewicz, Jacek Jemielity, Joanna Kowalska
Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Grant
Filed:
September 29, 2004
Date of Patent:
July 4, 2006
Assignee:
Becton Dickinson and Company
Inventors:
Zbigniew Darzynkiewicz, Frank Traganos, Gloria Juan, Stefan Gruenwald
Abstract: The invention provides assay methods and reagents useful for evaluating the level of enzyme activities within living cells. Enzyme activity levels within living cells, such as caspases and Serine proteases, can be key determinates in assessing; 1) the apoptotic state of a cell, 2) the presence of tumor (cancer) cells, 3) the predictive efficacy of a chemotherapeutic treatment regimen using a particular therapeutic agent or process, 4) the probability of graft rejection or acceptance, identification of the up or down regulation relationships of serine proteases and caspases within living cell systems, provides a rapid, yet finely tuned mechanism for predicting the current and future state of these cell populations, and 5) the disease state status of a cell.
Type:
Application
Filed:
June 21, 2004
Publication date:
June 23, 2005
Inventors:
David Phelps, Gary Johnson, Brian Lee, Zbigniew Darzynkiewicz, Jerzy Grabarek
Abstract: Methods, reagents, and kits are provided that permit flow cytometric determination of the phosphorylation status of retinoblastoma susceptibility gene protein (pRB) in individual cells. Methods are described that permit the hypophosphorylated, active, form of pRB to be measured either as an absolute quantity or as a proportion of total cellular pRB. Further described are methods that permit pRB phosphorylation status to be correlated with cell cycle phase and with protein components of the cell cycle. Screening of chemical compounds for antiproliferative and antineoplastic activity using the flow cytometric assays is demonstrated. Reagent kits that facilitate the subject methods are also provided.
Type:
Grant
Filed:
February 24, 1999
Date of Patent:
November 23, 2004
Assignee:
Becton, Dickinson and Company
Inventors:
Zbigniew Darzynkiewicz, Frank Traganos, Gloria Juan, Stefan Gruenwald
Abstract: The invention pertains to the field of DNA detection for basic research, medical diagnostic testing, and forensic testing. Methods are provided for end labeling of DNA strands without a denaturation step so that cellular morphology can be better preserved. The DNA strands are first incubated with a halogenated deoxynucleotide triphosphate, such as brominated deoxyuridine triphosphate (BrdUTP), and an enzyme which can catalyze the addition of the halogenated deoxynucleotide to the 3' OH ends of the DNA strand, such as terminal deoxynucleotidyl transferase (TdT). The resulting modified DNA strands are then incubated with a labeled antibody, such as a fluoresceinated monoclonal antibody, that binds specifically to the halogenated deoxynucleotide. The label is then detected, e.g., by flow cytometry. The methods have utility in detecting apotosis, DNA synthesis and/or repair, and as general methods for end labeling DNA.
Type:
Grant
Filed:
October 22, 1996
Date of Patent:
June 15, 1999
Inventors:
Zbigniew Darzynkiewicz, Xun Li, Frank Traganos
Abstract: The ability to synthesize capped RNA transcripts in vitro has been of considerable value in a variety of applications. However, one-third to one-half of the caps have, until now, been incorporated in the reverse orientation. Such reverse caps impair the translation of in vitro-synthesized mRNAs. Novel cap analogues, such as P1-3?-deoxy-7-methylguanosine-5? P3-guanosine-5? triphosphate and P1-3?-O,7-dimethylguanosine-5? P3-guanosine-5? triphosphate, have been designed that are incapable of being incorporated into RNA in the reverse orientation. Transcripts produced with SP6 polymerase using “anti-reverse” cap analogues were of the predicted length. Analysis of the transcripts indicated that reverse caps were not formed. The in vitro translational efficiency of transcripts with the novel “anti-reverse” cap analogues was significantly higher than that of transcripts formed with conventional caps.
Type:
Application
Filed:
July 10, 2006
Publication date:
November 9, 2006
Inventors:
Edward Darzynkiewicz, Robert Rhoads, Janusz Stepinski
Abstract: New RNA cap analogs are disclosed containing one or more phosphorothioates groups. The analogs also contain modifications at the 2?-O position of 7-methylguanosine that prevent them from being incorporated in the reverse orientation during in vitro synthesis of mRNA and that hence are “anti-reverse cap analogs” (ARCAs). The ARCA modification ensures that the S atom is precisely positioned within the active sites of cap-binding proteins in both the translational and decapping machinery. The new S-ARCA analogs are resistant to in vivo decapping enzymes. Some S-ARCAs have a higher affinity for eIF4E than the corresponding analogs not containing a phosphorothioate group. When mRNAs containing the various S-ARCAs are introduced into cultured cells, some are translated as much as five-fold more efficiently than mRNAs synthesized with the conventional analog m7GpppG.
Type:
Application
Filed:
June 19, 2008
Publication date:
September 16, 2010
Applicant:
BOARD OF SUPERVISORS OF LOUSIANA STATE UNIVERSITY
Inventors:
Jacek Jemielity, Ewa M. Grudzien-Nogalska, Joanna Kowalska, Edward Darzynkiewicz, Robert E. Rhoads