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发明公开
CN108018288A 用于突变ROSA26基因的系统及其应用 审中-实审

用于突变ROSA26基因的系统及其应用
摘要：
本发明公开了用于突变ROSA26基因的系统及其应用。本发明提供的用于突变ROSA26基因系统为CRISPR/Cas9系统，该系统包括靶向ROSA26基因的sgRNA，sgRNA的靶序列为如下A1)、A2)或A3)：A1)序列表中序列2的核苷酸序列；A2)与A1)限定的DNA序列具有75％或75％以上同一性的由A1)衍生的DNA序列；A3)在严格条件下与A1)限定的DNA序列杂交的由A1)衍生的DNA序列；该系统还包括Cas9和含有Sfat1基因表达盒的供体DNA。实验证明，利用本发明的系统可以成功敲除ROSA26基因，并定点整合Sfat1基因，最终成功得到表达Sfat1的动物。
摘要（英）：

The invention discloses a system for mutating ROSA26 genes and application thereof. The system for mutating the ROSA26 genes provided by the invention is a CRISPR/Cas9 system. The system includes an sgRNA targeting the ROSA26 genes. A target sequence of the sgRNA is the following sequence: A1), A2), or A3), wherein A1) refers to a nucleotide sequence of sequence 2; A2) refers to a DNA sequence derived from A1) and having 75% or above identity with the DNA sequence defined by A1); A3) refers to a DNA sequence derived from A1) and hybridized with the DNA sequence defined by A1) under stringent conditions. The system also comprises Cas9 and donor DNA containing a Sfat1 gene expression cassette. Experiments prove that the ROSA26 gene can be successfully knocked out by using the system, and theSfat1 gene can be integrated in a targeted manner. Finally, an animal expressing Sfat1 is successfully obtained.

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