The Uab Research Foundation (the University of Alabama at Birmingham)

University of Pennsylvania

University of Washington

Tim M Townes

Ralph L Brinster

Richard D Palmiter

Steven L McCune

Thomas M Ryan

Richard R Behringer

Chia-Wei Chang

David Donze

Lei Ding

Kevin M Pawlik

Provided herein are CRISPR/Cas9 complexes and method of using same.

CRISPR/Cas9 complex for genomic editing

Provided herein are CRIS-PR/Cas9 complexes and methods of using same.

CRISPR/Cas9 complex for introducing a functional polypeptide into cells of blood cell lineage

Methods of producing induced pluripotent stem (iPS) cells are provided. For example, a method of producing an iPS cell from a differentiated cell, which includes transforming the differentiated cell with a first vector comprising a nucleic acid sequence comprising a nucleic acid sequence encoding an Oct4, a nucleic acid sequence encoding a Sox2, and a nucleic acid sequence encoding a Klf4. Each of the nucleic acid sequences are separated from each other by a first and second viral 2A sequence. The method described can further comprise culturing the transformed cell under conditions that allow for the production of an iPS cell and isolating the cultured iPS cell.

Polycistronic vector for human induced pluripotent stem cell production

The present invention relates to the synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic non-human animals and erythroid cell lines. It is based on the discovery that two of the five hypersensitivity sites of the &bgr;-globin locus are sufficient to result in high level expression of human &agr;- or &bgr;-globin transgenes.

Synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic non-human mammals

The invention features &dgr;-erythroid kr&uuml;ppel-like factors (&dgr;-EKLFs), and methods of using nucleic acids encoding &dgr;-EKLFs to increase &dgr;-globin gene expression in a cell.

Method for identifying a &dgr;-eklf

Nucleic acid molecule encoding a &bgr;-erythroid kr&uuml;ppel-like factor that binds to a &dgr;-globin promoter

The present invention provides novel recombinant nucleic acid vectors which may be used to produce .alpha.-globin as well as other proteins of interest in quantity in the red blood cells of transgenic animals or cell cultures of erythroid lineage. The present invention also provides for the transgenic animals which contain these recombinant nucleic acid vectors. The vectors of the invention comprise at least one of the major DNase I hypersensitivity sites associated with the .beta.-globin locus together with a gene of interest. According to various embodiments of the invention, the vectors may be used to create transgenic animals or to transfect cells in culture. In a specific embodiment of the invention, a vector which comprises two DNase I hypersensitivity sites together with the human .alpha.-globin gene is used to create transgenic animals which produce human .alpha.-globin protein in erythroid tissues, including red blood cells. In a preferred specific embodiment of the invention, transgenic animals are created which comprise human .alpha.-globin and .beta.-globin genes, each under the transcriptional influence of two .beta.-globin locus DNase hypersensitivity sites; these transgenic animals express human hemoglobin in their erythroid tissues, and can be used to produce human hemoglobin in quantity.

Nucleic acid vectors comprising DNase I hypersensitive sites

Disclosed are anti-sickling human hemoglobins for use as sickle cell anemia therapeutics.

Anti-sickling hemoglobin

Transgenic, recombinantly cross-linked polymeric human hemoglobins suitable as cell-free blood substitutes have been produced. A plurality of DNA constructs have been designed for efficient expression of modified human hemoglobins in the erythrocytes of the non-human transgenic animals. Substantially pure, non-immunogenic, artificial human hemoglobins are then easily obtained from the erythroid cells of the transgenic animals.

Transgenic,cross-linked hemoglobin

The present invention relates to the synthesis of functional human hemoglobin and other proteins in erythroid tissues of transgenic non-human animals and erythroid cell lines. It is based on the discovery that two of the five hypersensitivity sites of the .beta.-globin locus are sufficient to result in high level expression of human .alpha.- or .beta.-globin transgenes. The present invention also provides for novel recombinant nucleic acid vectors which may be used to produce .alpha.-globin as well as other proteins of interest in quantity in the red blood cells of transgenic animals or cell cultures of erythroid lineage. The vectors of the invention comprise at least one of the major DNase I hypersensitivity sites associated with the .beta.-globin locus together with a gene of interest. In a specific embodiment of the invention, a vector which comprises two DNase I hypersensitivity sites together with the human .alpha.-globin gene is used to create transgenic animals which produce human .alpha.-globin protein in erythroid tissues, including red blood cells. In a preferred specific embodiment of the invention, transgenic animals are created which comprise human .alpha.-globin and .beta.-globin genes, each under the transcriptional influence of two .beta.-globin locus DNase hypersensitivity sites; these transgenic animals express human hemoglobin in their erythroid tissues, and can be used to produce human hemoglobin in quantity.

Growing community of inventors

Birmingham, AL, United States of America

Tim M Townes