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Rationally designed fluorogenic protease reporter visualizes spatiotemporal dynamics of apoptosis in vivo

Author:
To, Tsz-Leung, Piggott, Beverly J., Makhijani, Kalpana, Yu, Dan, Jan, Yuh Nung, Shu, Xiaokun
Source:
Proceedings of the National Academy of Sciences of the United States of America 2015 v.112 no.11 pp. 3338-3343
ISSN:
0027-8424
Subject:
apoptosis, biological development, drugs, engineering, enzyme activity, fluorescent proteins, image analysis, monitoring, proteinase inhibitors, proteinases, screening, signal transduction
Abstract:
Fluorescence resonance energy transfer-based reporters have been widely used in imaging cell signaling; however, their in vivo application has been handicapped because of poor signal. Although fluorogenic reporters overcome this problem, no such reporter of proteases has been demonstrated for in vivo imaging. Now we have redesigned an infrared fluorescent protein so that its chromophore incorporation is regulated by protease activity. Upon protease activation, the infrared fluorogenic protease reporter becomes fluorescent with no requirement of exogenous cofactor. To demonstrate biological applications, we have designed an infrared fluorogenic executioner-caspase reporter, which reveals spatiotemporal coordination between cell apoptosis and embryonic morphogenesis, as well as dynamics of apoptosis during tumorigenesis in Drosophila . The designed scaffold may be used to engineer reporters of other proteases with specific cleavage sequence.
Agid:
2303529