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. 2020 Jun 7;23(6):101248. doi: 10.1016/j.isci.2020.101248

Figure 2.

Figure 2

BATTLE-2 (with Shadow Fighter): Split-Tunable and Multi-sparse Allocation of Transgenes Using the Battle of the Recombinases

(A) In BATTLE-2 technology, Cre, FLPO, and Dre are expressed under the control of CamKIIa promoter. Then, Cre excises FLPO and Dre transgenes, whereas FLPO excises Cre and Dre transgenes. In addition, Dre excises FLPO and Cre transgenes. Using AAVs, Cre induces mCherry expression and FLPO induces YFP expression. However, Dre as a shadow fighter did not induce expression of transgenes.

(B) (Left) A representative maximum intensity projection image of dentate gyrus infected with BATTLE-2 system when the ratio of Dre:Cre:FLPO was 1:1:1. (Right) Quantification of cell number of neurons expressing fluorescent protein in the dentate gyrus. Data are represented as mean ± SEM.

(C) (Left) Representative maximum intensity projection image of dentate gyrus infected with BATTLE-2 system when the ratio of Dre:Cre:FLPO was 200:1:10. (Right) Quantification of cell number of neurons expressing fluorescent protein in the dentate gyrus. Data are represented as mean ± SEM.

(D) Representative maximum intensity projection image of dentate gyrus infected with conventional virus AAV-CAG-GFP. Note that the morphologies of mossy cells are unclear.

(E) Representative maximum intensity projection image of dentate gyrus infected with BATTLE-2 (double sparse). Note that the morphologies of multiple mossy cells are clear using BATTLE-2 double sparse expression.