DR. LLOYD G MITCHELL, MD
Medical Practice at Gretna St, Bethesda, MD
License number
Maryland D0032764
Category
Medical Practice
Type
Molecular Genetic Pathology
Address
Address
4519 Gretna St, Bethesda, MD 20814
Phone
(240) 597-1967
Professional information
Transgenic Animal Model For Spliceosome-Mediated Rna Trans-Splicing
US Patent:
2003020, Oct 30, 2003
Filed:
Apr 30, 2002
Appl. No.:
10/136723
Inventors:
Madaiah Puttaraju - Germantown MD, US
Lloyd Mitchell - Bethesda MD, US
John Engelhardt - Iowa City IA, US
Xiaoming Liu - Iowa City IA, US
Lloyd Mitchell - Bethesda MD, US
John Engelhardt - Iowa City IA, US
Xiaoming Liu - Iowa City IA, US
International Classification:
A01K067/027
US Classification:
800/018000
Abstract:
The present invention relates to development of an animal model system for in vivo testing of spliceosome-mediated RNA trans-splicing reactions. The present invention provides transgenic animals, and methods for generating such animals, that have been genetically engineered to expresses a target precursor messenger RNA molecule (target pre-mRNA) that serves as a substrate for a trans-splicing reaction. Specifically, the transgenic animals contain at least one transgene capable of expressing a target pre-mRNA molecule. The invention provides methods, based on utilization of the transgenic animals, for assessing the specificity and efficiency of a pre-trans-splicing molecule (PTM) designed to interact with a target pre-mRNA and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule. The present invention further relates to the transgenic expression of PTM molecules in animals to determine gene function, i.e, functional genetics. The present invention is based on the successful generation of a transgenic animal expressing a target pre-mRNA and, moreover, the use of that animal to detect accurate in vivo trans-splicing reactions in the presence of a PTM.
Dr. Lloyd G Mitchell, Bethesda MD - MD (Doctor of Medicine)
Specialties:
Molecular Genetic Pathology
Address:
4519 Gretna St, Bethesda 20814
(240) 597-1967 (Phone)
(240) 597-1967 (Phone)
Languages:
English
Education:
Medical School
University of Maryland At Baltimore / Professional Schools
Graduated: 1984
University of Maryland At Baltimore / Professional Schools
Graduated: 1984
Spliceosome Mediated Rna Trans-Splicing For Correction Of Skin Disorders
US Patent:
2004024, Dec 9, 2004
Filed:
Jul 17, 2003
Appl. No.:
10/621867
Inventors:
Lloyd Mitchell - Bethesda MD, US
Madaiah Puttaraju - Germantown MD, US
Guenter Dallinger - Linz, AT
Alfred Klausegger - Salzburg, AT
Johann Bauer - Salzburg, AT
Madaiah Puttaraju - Germantown MD, US
Guenter Dallinger - Linz, AT
Alfred Klausegger - Salzburg, AT
Johann Bauer - Salzburg, AT
International Classification:
C12Q001/68, C07H021/04, A61K048/00, C12N005/08
US Classification:
435/006000, 435/069100, 435/320100, 435/371000, 536/023500
Abstract:
The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated RNA trans-splicing. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA). In particular, the PTMs of the present invention can be genetically engineered to interact with a specific target pre-mRNA expressed in cells of the skin so as to result in correction of genetic defects responsible for a variety of different skin disorders to encode a reporter molecule or protein that may have therapeutic benefit. The compositions of the invention further include recombinant vectors systems capable of expressing the PTMs of the invention and cells expressing said PTMs. The methods of the invention encompass contacting the PTMs of the invention with specific target pre-mRNA expressed within cells of the skin under conditions in which a portion of the PTM is trans-spliced to a portion of the target pre-mRNA to form a chimeric RNA molecule wherein the genetic defect in the specific gene has been corrected. The present invention is based on the successful trans-splicing of the collagen XVII pre-mRNA thereby establishing the usefulness of trans-splicing for correction of skin specific genetic defects. The methods and compositions of the present invention can be used in gene therapy for treatment of specific disorders of the skin, i.e., genodermatoses, such as epidermal fragility disorders, keratinization disorders, hair disorders and pigmentation disorders as well as cancers of the skin.
Correction Of Alpha-1-Antitrypsin Genetic Defects Using Spliceosome Mediated Rna Trans Splicing
US Patent:
8053232, Nov 8, 2011
Filed:
Jan 21, 2005
Appl. No.:
11/040634
Inventors:
Madaiah Puttaraju - Germantown MD, US
Edward Otto - Reston VA, US
Mariano A. Garcia-Blanco - Durham NC, US
Gerard J. McGarrity - Gaithersburgh MD, US
Gary F. Temple - Washington Grove MD, US
Lloyd G. Mitchell - Bethesda MD, US
Colette Cote - Gaithersburgh MD, US
S. Gary Mansfield - Montgomery Village MD, US
Edward Otto - Reston VA, US
Mariano A. Garcia-Blanco - Durham NC, US
Gerard J. McGarrity - Gaithersburgh MD, US
Gary F. Temple - Washington Grove MD, US
Lloyd G. Mitchell - Bethesda MD, US
Colette Cote - Gaithersburgh MD, US
S. Gary Mansfield - Montgomery Village MD, US
Assignee:
VIRxSYS Corporation - Gaithersburg MD
International Classification:
C12N 5/00, C12N 15/12, C12N 15/63, C12N 15/86
US Classification:
435325, 4353201
Abstract:
The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated RNA trans-splicing. The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a SERPINA1 target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA). In particular, the PTMs of the present invention include those genetically engineered to interact with SERPINA1 target pre-mRNA so as to result in correction of SERPINA1 genetic defects responsible for AAT deficiency. The PTMs of the invention may also comprise sequences that are processed out of the PTM to yield duplex siRNA molecules directed specifically to mutant SERPIN A1 mRNAs. Such duplexed siRNAs are designed to reduce the accumulation of toxic AAT protein in liver cells. The methods and compositions of the present invention can be used in gene therapy for correction of SERPINA1 disorders such as AAT deficiency.
Screening Method For Identification Of Efficient Pre-Trans-Splicing Molecules
US Patent:
2006015, Jul 13, 2006
Filed:
Jul 26, 2004
Appl. No.:
10/898748
Inventors:
Lloyd Mitchell - Bethesda MD, US
Madaiah Puttaraju - Germantown MD, US
Mariano Garcia-Blanco - Durham NC, US
Edward Otto - Reston VA, US
Yanping Yang - Rockville MD, US
Madaiah Puttaraju - Germantown MD, US
Mariano Garcia-Blanco - Durham NC, US
Edward Otto - Reston VA, US
Yanping Yang - Rockville MD, US
International Classification:
C40B 40/10, C40B 40/08
US Classification:
435006000, 435007100
Abstract:
The present invention provides methods and compositions for rapid high capacity functional screening to identify optimal pre-trans-splicing molecules (PTMs). The compositions of the invention include PTM expression libraries capable of encoding candidate PTMs designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA). The candidate PTMs of the invention encode a portion of a first reporter molecule and may encode one or more other reporter molecules, which can be used to select for cells expressing optimal PTMs (efficient and specific). The compositions of the invention also include cells that express a target pre-mRNA encoding the remaining portion of the first reporter molecule. The screening methods of the invention encompass (i) contacting a PTM expression library with cells expressing a target pre-mRNA under conditions in which a trans-splicing reaction will occur in the presence of an optimal PTM (expressed by the library vector) resulting in the formation of a chimeric repaired RNA molecule capable of encoding at least one reporter molecule; (ii) selecting for cells expressing the repaired reporter molecule wherein expression of the reporter molecule indicates the presence of an optimal PTM in the selected cell; and (iii) identifying the optimal PTM expressed in the selected cell(s). The additional reporter molecule(s) can be used to assess both specific and non-specific trans-splicing, as well direct PTM expression.
Trans-Splicing Mediated Photodynamic Therapy
US Patent:
7399753, Jul 15, 2008
Filed:
Sep 9, 2003
Appl. No.:
10/658617
Inventors:
Lloyd G. Mitchell - Bethesda MD, US
Edward Otto - Great Falls VA, US
Carl R. Merril - Bethesda MD, US
Edward Otto - Great Falls VA, US
Carl R. Merril - Bethesda MD, US
Assignee:
Virxsys Corporation - Gaithersburg MD
International Classification:
C12N 5/10, C12N 15/87, C12N 15/63, C12N 15/66, C12N 15/00, A61K 31/711, C12N 15/861
US Classification:
514 44, 435325, 435455, 435 691, 435 911, 435 913
Abstract:
The present invention provides methods and compositions for conferring selective death on cells expressing a specific target precursor messenger RNA (selective target pre-mRNA). The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) expressed within a cell and mediate a trans-splicing reaction resulting in the generation of a novel chimeric mRNA molecule (chimeric mRNA) capable of encoding a light producing protein or enzyme. Cell death is further mediated by the presence of a photosensitizer which upon photoactivation produces cytotoxicity.
Sequencing Dna; A Modification Of The Polymerase Chain Reaction
US Patent:
5817797, Oct 6, 1998
Filed:
Jun 1, 1988
Appl. No.:
7/200876
Inventors:
Lloyd G. Mitchell - Bethesda MD
Carl R. Merril - Rockville MD
Carl R. Merril - Rockville MD
Assignee:
The United States of America as represented by the Department of Health
and Human Services - Washington DC
and Human Services - Washington DC
International Classification:
C07H 2100, C12Q 168, C12P 1934
US Classification:
536 253
Abstract:
The invention provides a process wherein a biotinylated oligonucleotide primer and an oligonucleotide primer which has not undergone biotinylation are used when amplifying a DNA sequence to facilitate separation of the DNA strands following the polymerase chain reaction process. The biotinylation/PCR product is then exposed to a support which will selectively bind the biotinylated strand to allow selective elution of the product.
Methods And Compositions For Determining The Purity Of And Purifying Chemically Synthesized Nucleic Acids
US Patent:
8076461, Dec 13, 2011
Filed:
Dec 24, 2003
Appl. No.:
10/746430
Inventors:
Christopher D. J. Pearce - Leatherhead, GB
Lloyd G. Mitchell - Bethesda MD, US
Lloyd G. Mitchell - Bethesda MD, US
Assignee:
Proteome Sciences PLC - Surrey
International Classification:
C07H 21/02, C07H 21/04
US Classification:
53038821, 435 61, 435 71, 536 231
Abstract:
An antibody microarray is described comprising a plurality of antibodies immobilized on a substrate, wherein each antibody specifically binds to a synthetic oligomer (e. g. , an oligonucleotide or oligopeptide) having an organic protecting group covalently bound thereto, which antibody does not bind to that synthetic oligomer when the organic protecting group is not covalently bound thereto. Methods of making and using such antibodies are disclosed, along with cells for making such antibodies. Methods of making and using such antibody microarrays are also disclosed.
Methods And Compositions For Use In Spliceosome Mediated Rna Trans-Splicing
US Patent:
2003015, Aug 14, 2003
Filed:
Feb 12, 2002
Appl. No.:
10/075028
Inventors:
Lloyd Mitchell - Bethesda MD, US
Mariano Garcia-Blanco - Durham NC, US
Madaiah Puttaraju - Germantown MD, US
Mariano Garcia-Blanco - Durham NC, US
Madaiah Puttaraju - Germantown MD, US
International Classification:
C12P019/34, C12N005/00
US Classification:
435/091200, 435/375000
Abstract:
The present invention provides methods and compositions for delivery of synthetic pre-trans-splicing molecules (synthetic PTMs) into a target cell. The compositions of the invention include synthetic pre-trans-splicing molecules (PTMs) with enhanced stability against chemical and enzymatic degradation. The synthetic PTMs are designed to interact with a natural target precursor messenger RNA molecule (target pre-mRNA) and mediate a trans-splicing reaction resulting in the generation of a novel chimeric RNA molecule (chimeric RNA).
Trans-Splicing Mediated Photodynamic Therapy
US Patent:
2009020, Aug 13, 2009
Filed:
Jun 9, 2008
Appl. No.:
12/135247
Inventors:
Lloyd G. Mitchell - Bethesda MD, US
Edward Otto - Great Falls VA, US
Carl R. Merril - Bethesda MD, US
Edward Otto - Great Falls VA, US
Carl R. Merril - Bethesda MD, US
Assignee:
VIRXSYS CORPORATION - Gaithersburg MD
International Classification:
C12N 15/87, C12N 15/11, C12N 1/21
US Classification:
435471, 536 232, 43525233
Abstract:
The present invention provides methods and compositions for conferring selective death on cells expressing a specific target precursor messenger RNA (selective target pre-mRNA). The compositions of the invention include pre-trans-splicing molecules (PTMs) designed to interact with a target precursor messenger RNA molecule (target pre-mRNA) expressed within a cell and mediate a trans-splicing reaction resulting in the generation of a novel chimeric mRNA molecule (chimeric mRNA) capable of encoding a light producing protein or enzyme. Cell death is further mediated by the presence of a photosensitizer which upon photoactivation produces cytotoxicity.