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. 2022 May 16;38(11):1383–1396. doi: 10.1007/s12264-022-00860-7

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — A dual-AAV system for sparse labeling of cell-type-specific neurons. Schematic of the design of the dual-AAV system for sparse labeling, comprising a “controller” and an “amplifier” AAV vector (upper left). When these two vectors are mixed and injected into the Cre-expressing mouse, leakage of TRE drives the weak expression of Flp in a few Cre⁺ neurons. Flp subsequently flips the GFP-IRES-tTA cassettes in the amplifier and drives the low level of GFP and tTA expression in these sparsely-labeled neurons (right panel). The small amount of tTA further binds to the TRE and potentiates the expression of Flp and GFP (lower middle), thereby triggering positive feedback to enhance GFP expression in sparse neurons (lower left). TRE, tetracycline response element; Flp, flippase; IRES, internal ribosome entry site; tTA, tetracycline trans-activator.