Lloyd Earnest Mitchell

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

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.

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

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.

US Patent:

20020193580, Dec 19, 2002

Filed:

Feb 12, 2002

Appl. No.:

10/076248

Inventors:

Lloyd Mitchell - Bethesda MD, US
Mariano Garcia-Blanco - Durham NC, US
Madaiah Puttaraju - Germantown MD, US

International Classification:

C07H021/02
C07H021/04

US Classification:

536/023100

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).

US Patent:

20030153054, 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

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).

US Patent:

20040018622, Jan 29, 2004

Filed:

Jul 17, 2002

Appl. No.:

10/198447

Inventors:

Lloyd Mitchell - Bethesda MD, US
Madaiah Puttaraju - Germantown MD, US
Guenter Dallinger - Linz, AT
Alfred Klausegger - Salzburg, AT
Johann Bauer - Salzburg, AT

International Classification:

A61K048/00
C12N005/08

US Classification:

435/371000, 514/044000, 424/093210

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 are 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. 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.

US Patent:

20040038396, Feb 26, 2004

Filed:

Jun 5, 2002

Appl. No.:

10/360787

Inventors:

Lloyd Mitchell - Bethesda MD, US
S. Mansfield - Montgomery Village MD, US

International Classification:

C12N005/06

US Classification:

435/325000

Abstract:

The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated 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 are genetically engineered to interact with factor VIII (FVIII) target pre-mRNA so as to result in correction of clotting FVIII genetic defects responsible for hemophilia A. The compositions of the invention further include recombinant vector 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 a FVIII target pre-mRNA under conditions in which a portion of the PTM is trans-spliced to a portion of the target pre-mRNA to form a RNA molecule wherein the genetic defect in the FVIII gene has been corrected. The methods and compositions of the present invention can be used in gene therapy for correction of FVIII disorders such as hemophilia A.

US Patent:

20040058344, Mar 25, 2004

Filed:

Feb 25, 2003

Appl. No.:

10/374784

Inventors:

Lloyd Mitchell - Bethesda MD, US
M. Puttaraju - Germantown MD, US
S. Mansfield - Montogomery Village MD, US

International Classification:

C12Q001/68
C12P021/02
C12N005/06
C12N015/85

US Classification:

435/006000, 435/069100, 435/455000, 435/320100, 435/325000

Abstract:

The present invention provides methods and compositions for imaging of gene expression in cells. 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 RNA molecule (chimeric RNA) capable of encoding a reporter molecule. The PTMs of the invention are designed to interact with target pre-mRNAs thereby providing a method for detection of target pre-mRNA expression. The methods and compositions of the invention may be utilized to monitor the expression of specific genes within a cell. In instances where specific gene expression is associated with a disease, the present invention provides diagnostic methods and compositions. Such diseases include infectious diseases, proliferative disorders such as cancer, genetic, neurological and metabolic disorders, to name a few. Additionally, the present invention may be used in screening assays to identify compounds capable of modulating gene expression or in assays designed to identify protein/protein interactions. The invention is demonstrated by way of example in which papilloma virus gene expression within a cell was detected using a bioluminescence assay system.

US Patent:

20040180429, Sep 16, 2004

Filed:

Jun 5, 2003

Appl. No.:

10/456273

Inventors:

Lloyd Mitchell - Bethesda MD, US
John Englehardt - Iowa City IA, US
Xiao Liu - Iowa City IA, US

International Classification:

C12N005/08

US Classification:

435/366000, 435/320100

Abstract:

The present invention provides methods and compositions for generating novel nucleic acid molecules through targeted spliceosomal mediated trans-splicing in stem cells. The compositions of the invention include stem cells engineered to express 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 novel chimeric RNA molecules (chimeric RNA). In particular, the stem cells of the present invention are genetically engineered to express a PTM that will interact with a specific target pre-mRNA expressed within a stem cell as it differentiates so as to result in correction of a genetic defect responsible for a genetic disorder. The methods of the invention encompass transferring a nucleic acid molecule capable of encoding a PTM of interest into a stem cell followed by transplantation of the PTM modified stem cell into a host. As the stem cell differentiates the target pre-mRNA is expressed thereby providing the substrate for a trans-splicing reaction. The present invention is based on the successful transfer and expression of a nucleic acid molecule encoding a PTM capable of interacting with a cystic fibrosis transmembrane conductance regulator (CFTR) pre-mRNA into primary human surface airway progenitor cells. The methods and compositions of the present invention can be used to correct genetic defects associated with a variety of different disorders such as cystic fibrosis, hemophilia, sickle cell anemia, Tay-Sachs disease, thalassemias, polycystic kidney disease and muscular dystrophy, to name a few.