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.
MeSH terms
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Animals
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Base Sequence
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CD2 Antigens / genetics*
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CD2 Antigens / immunology
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CD4-Positive T-Lymphocytes / cytology
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CD4-Positive T-Lymphocytes / immunology
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CD8-Positive T-Lymphocytes / cytology
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CD8-Positive T-Lymphocytes / immunology
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CRISPR-Cas Systems*
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Gene Editing / methods*
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Gene Silencing*
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Genetic Engineering
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Genetic Vectors / chemistry
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Genetic Vectors / metabolism
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Immunophenotyping
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Lymph Nodes / cytology
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Lymph Nodes / immunology
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Mice
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Mice, Transgenic
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Mutation
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Promoter Regions, Genetic
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RNA, Guide, CRISPR-Cas Systems / genetics*
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RNA, Guide, CRISPR-Cas Systems / metabolism
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Spleen / cytology
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Spleen / immunology
Substances
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CD2 Antigens
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RNA, Guide, CRISPR-Cas Systems