Helmholtz Zentrum Munchen Deutsches Forschungszentrum Fur Gesundheit Und Umwelt

Other

Max-planck-gesellschaft Zur F&#xf6;rderung Der Wissenschaften E. V.

Gsf-forschungszentrum Fur Umwelt Und Gesundheit, Gmbh

Frankgen Biotechnologie Ag

Klinikum Rechts Der Isar Der Technischen Universitat München

Wolfgang Wurst

Ralf Kühn

Oskar Ortiz Sanchez

Alain Prochiantz

Frank Schnütgen

Ralf Kuehn

Harald Von Melchner

Yuanyuan Chu

Thomas Floss

Patricia Ruiz

Melanie Meyer

Jens Hansen

Dong-Jiunn Jeffery Truong

Gil Gregor Westmeyer

Patricia Steuber-Buchberger

Silke De-Zolt

Ralf KÜHN

The present invention relates to a method of producing a non-human, mammalian oocyte carrying a modified target sequence in its genome, the method comprising the steps of introducing into a non-human, mammalian oocyte: (a) a clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated protein 9 (Cas9 protein) or a nucleic acid molecule encoding said Cas9 protein; and (b-i) a target sequence specific CRISPR RNA (crRNA) and a trans-activating crRNA (tracr RNA) or a nucleic acid molecule encoding said RNAs; or (b-ii) a chimaeric RNA sequence comprising a target sequence specific crRNA and tracrRNA or a nucleic acid molecule encoding said RNA; wherein the Cas9 protein introduced in (a) and the RNA sequence(s) introduced in (b-i) or (b-ii) form a protein/RNA complex that specifically binds to the target sequence and introduces a single or double strand break within the target sequence. The present invention further relates to the method of the invention, wherein the target sequence is modified by homologous recombination with a donor nucleic acid sequence further comprising the step: (c) introducing a nucleic acid molecule into the cell, wherein the nucleic acid molecule comprises the donor nucleic acid sequence and regions homologous to the target sequence. The present invention also relates to a method of producing a non-human mammal carrying a modified target sequence in its genome.

Gene editing in the oocyte by Cas9 nucleases

Gene editing in the oocyte by CAS9 nucleases

The present invention relates to a novel class of gene trap vector (enhanced gene trap vectors, eGTV) for efficiently identifying silent or weakly expressed target genes in mammalian genomes, methods of their production and methods for identifying and mutating target genes by using the enhanced gene trap vectors. The gene trap vectors of the present invention can also be used for inducing the expression of silent genes and enhancing the expression of weakly expressed genes. The use of the enhanced gene trap vectors for creating transgenic organisms to identify gene function and to validate pharmaceutical compounds prior to clinical applications is a further aspect of the present invention.

Enhancer-containing gene trap vectors for random and targeted gene trapping

The present invention relates to a fusion protein comprising a Caspase domain or a functionally active variant thereof and a ligand binding domain of a nuclear hormone receptor, a nucleic acid coding for the fusion protein, a vector or cell comprising the nucleic acid, a method of producing the fusion protein, a non-human transgenic animal containing the nucleic acid, the use of the fusion protein for ligand-mediated induction of apoptosis of a cell, or for studying the function of a cell, tissue and/or organ or the use of a transgenic organism for studying the function of a cell at various developmental stages or as a disease model, a method for inducing apoptosis of a cell expressing a fusion protein or for identifying a ligand, or a medicament comprising a fusion protein, the nucleic acid, the vector or the cell, particularly for the treatment of cancer or for or after transplantation, particularly as safety mechanism.

Fusion protein comprising a Caspase domain and a nuclear hormone receptor binding domain and methods and uses thereof

Transcription factor responsive elements, target genes, and co-factors of a transcription factor, either individually or in combination, are identified by integrating into the genome of a eukaryotic cell a gene trap vector that includes a reporter gene, a polyadenylation site, and a selectable marker gene, selecting the cells in which the gene trap vector is successfully integrated and contacting those cells with the transcription factor, then identifying and cultivating those cells having a gene into which the gene trap vector has been integrated and in which the reporter gene activity has been altered by interaction between the transcription factor and a transcription factor responsive element of that gene, and identifying the transcription factor responsive element, target gene, or cofactor that interacts with the transcription factor.

Method for identification of target genes of transcription factors

Growing community of inventors

Munich, Germany

Wolfgang Wurst