Generation of transgenic mice expressing a FRET biosensor, SMART, that responds to necroptosis.
Shin MuraiKanako TakakuraKenta SumiyamaKenta MoriwakiKenta TeraiSachiko Komazawa-SakonTakao SekiYoshifumi YamaguchiTetuo MikamiKimi ArakiMasaki OhmurayaMichiyuki MatsudaHiroyasu NakanoPublished in: Communications biology (2022)
Necroptosis is a regulated form of cell death involved in various pathological conditions, including ischemic reperfusion injuries, virus infections, and drug-induced tissue injuries. However, it is not fully understood when and where necroptosis occurs in vivo. We previously generated a Forster resonance energy transfer (FRET) biosensor, termed SMART (the sensor for MLKL activation by RIPK3 based on FRET), which monitors conformational changes of MLKL along with progression of necroptosis in human and murine cell lines in vitro. Here, we generate transgenic (Tg) mice that express the SMART biosensor in various tissues. The FRET ratio is increased in necroptosis, but not apoptosis or pyroptosis, in primary cells. Moreover, the FRET signals are elevated in renal tubular cells of cisplatin-treated SMART Tg mice compared to untreated SMART Tg mice. Together, SMART Tg mice may provide a valuable tool for monitoring necroptosis in different types of cells in vitro and in vivo.
Keyphrases
- energy transfer
- cell cycle arrest
- cell death
- single molecule
- quantum dots
- induced apoptosis
- drug induced
- living cells
- fluorescent probe
- high fat diet induced
- endoplasmic reticulum stress
- sensitive detection
- liver injury
- gold nanoparticles
- pi k akt
- heart failure
- gene expression
- endothelial cells
- transcription factor
- wild type
- molecular dynamics
- label free
- cerebral ischemia
- coronary artery disease
- adipose tissue
- skeletal muscle
- acute coronary syndrome
- ischemia reperfusion injury
- brain injury
- newly diagnosed
- adverse drug