Protocol for highly selective transgene expression through the flip-excision switch system by using a unilateral spacer sequence in rodents

Autor: Natsuki Matsushita, Shigeki Kato, Kayo Nishizawa, Masateru Sugawara, Kosei Takeuchi, Yoshiki Miyasaka, Tomoji Mashimo, Kazuto Kobayashi
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 4, Pp 102667- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2023.102667
Popis: Summary: We present a protocol to induce Cre-dependent transgene expression in specific cell types in the rat brain, suppressing a leak expression in off-target cells, by using a flip-excision switch system with a unilateral spacer sequence. We describe steps for construction of transfer plasmids, preparation of adeno-associated viral vectors, intracranial injection, and detection of transgene expression. Our protocol provides a useful strategy for a better understanding of the structure and function of specific cell types in the complex neural circuit.For complete details on the use and execution of this protocol, please refer to Matsushita et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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