Journal
COMMUNICATIONS BIOLOGY
Volume 5, Issue 1, Pages -Publisher
NATURE PORTFOLIO
DOI: 10.1038/s42003-022-03944-2
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Funding
- KAKENHI grants [16K08527, 16H01488, 17H06061, 19K06910, 22K06487, 17H04026, 17K19636, 25118008]
- Smoking Research Foundation
- Japan Prize Foundation
- program for Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) from Japan Agency for Medical Research and Development, AMED [JP19dm0207057]
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Transgenic animals expressing fluorescent proteins are commonly used to label specific cells and proteins. In this study, a new Cre recombinase dependent on red fluorescent protein (RFP) was created, allowing selective targeting and tracing of RFP-dependent gene expression in the mouse and rat brain.
Transgenic animals expressing fluorescent proteins are widely used to label specific cells and proteins. By using a split Cre recombinase fused with mCherry-binding nanobodies or designed ankyrin repeat proteins, we created Cre recombinase dependent on red fluorescent protein (RFP) (Cre-DOR). Functional binding units for monomeric RFPs are different from those for polymeric RFPs. We confirmed selective target RFP-dependent gene expression in the mouse cerebral cortex using stereotaxic injection of adeno-associated virus vectors. In estrogen receptor-beta (Esr2)-mRFP1 mice and gastrin-releasing peptide receptor (Grpr)-mRFP1 rats, we confirmed that Cre-DOR can be used for selective tracing of the neural projection from RFP-expressing specific neurons. Cellular localization of RFPs affects recombination efficiency of Cre-DOR, and light and chemical-induced nuclear translocation of an RFP-fused protein can modulate Cre-DOR efficiency. Our results provide a method for manipulating gene expression in specific cells expressing RFPs and expand the repertory of nanobody-based genetic tools. A Cre recombinase dependent on red fluorescent protein (RFP) is generated, which expands the repertory of nanobody-based genetic tools by enabling selective targeting of RFP-dependent gene expression in the mouse and rat brain.
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