通过CRISPR/Cas9对小鼠CD2进行T细胞特异性失活。
T cell-specific inactivation of mouse CD2 by CRISPR/Cas9.
作者信息
Beil-Wagner Jane, Dössinger Georg, Schober Kilian, vom Berg Johannes, Tresch Achim, Grandl Martina, Palle Pushpalatha, Mair Florian, Gerhard Markus, Becher Burkhard, Busch Dirk H, Buch Thorsten
机构信息
Institute for Medical Microbiology, Immunology and Hygiene, Technische Universität München, Germany.
Institute of Laboratory Animal Science, University of Zurich, Schlieren, Switzerland.
出版信息
Sci Rep. 2016 Feb 23;6:21377. doi: 10.1038/srep21377.
The CRISPR/Cas9 system can be used to mutate target sequences by introduction of double-strand breaks followed by imprecise repair. To test its use for conditional gene editing we generated mice transgenic for CD4 promoter-driven Cas9 combined with guide RNA targeting CD2. We found that within CD4(+) and CD8(+) lymphocytes from lymph nodes and spleen 1% and 0.6% were not expressing CD2, respectively. T cells lacking CD2 carryied mutations, which confirmed that Cas9 driven by cell-type specific promoters can edit genes in the mouse and may thus allow targeted studies of gene function in vivo.
CRISPR/Cas9系统可通过引入双链断裂并随后进行不精确修复来突变靶序列。为了测试其在条件性基因编辑中的应用，我们构建了转基因小鼠，其表达由CD4启动子驱动的Cas9，并结合靶向CD2的引导RNA。我们发现，在来自淋巴结和脾脏的CD4(+)和CD8(+)淋巴细胞中，分别有1%和0.6%不表达CD2。缺乏CD2的T细胞携带突变，这证实了由细胞类型特异性启动子驱动的Cas9可以在小鼠体内编辑基因，从而可能允许在体内对基因功能进行靶向研究。
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