1. T cell-specific inactivation of mouse CD2 by CRISPR/Cas9.
- Author
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Beil-Wagner J, Dössinger G, Schober K, vom Berg J, Tresch A, Grandl M, Palle P, Mair F, Gerhard M, Becher B, Busch DH, and Buch T
- Subjects
- Animals, Base Sequence, CD2 Antigens immunology, CD4-Positive T-Lymphocytes cytology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes cytology, CD8-Positive T-Lymphocytes immunology, Genetic Engineering, Genetic Vectors chemistry, Genetic Vectors metabolism, Immunophenotyping, Lymph Nodes cytology, Lymph Nodes immunology, Mice, Mice, Transgenic, Mutation, Promoter Regions, Genetic, RNA, Guide, CRISPR-Cas Systems metabolism, Spleen cytology, Spleen immunology, CD2 Antigens genetics, CRISPR-Cas Systems, Gene Editing methods, Gene Silencing, RNA, Guide, CRISPR-Cas Systems genetics
- Abstract
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.
- Published
- 2016
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