发明名称 |
GENE EDITING IN THE OOCYTE BY CAS9 NUCLEASES |
摘要 |
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. |
申请公布号 |
WO2014131833(A1) |
申请公布日期 |
2014.09.04 |
申请号 |
WO2014EP53840 |
申请日期 |
2014.02.27 |
申请人 |
HELMHOLTZ ZENTRUM MÜNCHEN DEUTSCHES FORSCHUNGSZENTRUM FÜR GESUNDHEIT UND UMWELT (GMBH) |
发明人 |
KÜHN, RALF;WURST, WOLFGANG;ORTIZ SANCHEZ, OSKAR |
分类号 |
A01K67/027;C12N9/22;C12N15/89;C12N15/90 |
主分类号 |
A01K67/027 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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