Measurement of Secreted Embryonic Alkaline Phosphatase

Wang, Meiyan; Wang, Xinyi; Ye, Haifeng

doi:10.21769/bioprotoc.4600

Cited by 1 publication

(1 citation statement)

References 8 publications

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“…A600162, Sangon Biotech), and 20 µL of substrate solution containing 120 mM p-nitrophenyl phosphate (catalog no. 333338-18-4, Sangon Biotech)] were added to 80 µL of heat-inactivated (65 °C, 30 min) cell culture supernatant 69 . The time course of absorbance at 405 nm was measured at 37 °C using a Synergy H1 hybrid multimode microplate reader (BioTek Instruments) with Gen5 software (version 2.04).…”

Section: Methodsmentioning

confidence: 99%

AAV-delivered muscone-induced transgene system for treating chronic diseases in mice via inhalation

Wu,

Yu,

Wang

et al. 2024

Nat Commun

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Gene therapies provide treatment options for many diseases, but the safe and long-term control of therapeutic transgene expression remains a primary issue for clinical applications. Here, we develop a muscone-induced transgene system packaged into adeno-associated virus (AAV) vectors (AAVMUSE) based on a G protein-coupled murine olfactory receptor (MOR215-1) and a synthetic cAMP-responsive promoter (PCRE). Upon exposure to the trigger, muscone binds to MOR215-1 and activates the cAMP signaling pathway to initiate transgene expression. AAVMUSE enables remote, muscone dose- and exposure-time-dependent control of luciferase expression in the livers or lungs of mice for at least 20 weeks. Moreover, we apply this AAVMUSE to treat two chronic inflammatory diseases: nonalcoholic fatty liver disease (NAFLD) and allergic asthma, showing that inhalation of muscone—after only one injection of AAVMUSE—can achieve long-term controllable expression of therapeutic proteins (ΔhFGF21 or ΔmIL-4). Our odorant-molecule-controlled system can advance gene-based precision therapies for human diseases.

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Section: Methodsmentioning

confidence: 99%