This GFP-UtrCH probe was published earlier than the 17aa peptide LifeAct (PMID: 18536722). Someone told me that it is better than LifeAct. I have not tested it myself. It would be interesting to compare LifeAct with UtrCH. We know that LifeAct cannot label certain actin stress fibers (PMID: 19404250). What is the limitation of UtrCH?
The size of the domain is about 161aa, which is OK size as a probe. The sequences are
>gi|110611227:93-875 Homo sapiens utrophin (UTRN), mRNA
ATGGCCAAGTATGGAGAACATGAAGCCAGTCCTGACAATGGGCAGAACGAATTCAGTGATATCATTAAGT
CCAGATCTGATGAACACAATGACGTACAGAAGAAAACCTTTACCAAATGGATAAATGCTCGATTTTCAAA
GAGTGGGAAACCACCCATCAATGATATGTTCACAGACCTCAAAGATGGAAGGAAGCTATTGGATCTTCTA
GAAGGCCTCACAGGAACATCACTGCCAAAGGAACGTGGTTCCACAAGGGTACATGCCTTAAATAACGTCA
ACAGAGTGCTGCAGGTTTTACATCAGAACAATGTGGAATTAGTGAATATAGGGGGAACTGACATTGTGGA
TGGAAATCACAAACTGACTTTGGGGTTACTTTGGAGCATCATTTTGCACTGGCAGGTGAAAGATGTCATG
AAGGATGTCATGTCGGACCTGCAGCAGACGAACAGTGAGAAGATCCTGCTCAGCTGGGTGCGTCAGACCA
CCAGGCCCTACAGCCAAGTCAACGTCCTCAACTTCACCACCAGCTGGACAGATGGACTCGCCTTTAATGC
TGTCCTCCACCGACATAAACCTGATCTCTTCAGCTGGGATAAAGTTGTCAAAATGTCACCAATTGAGAGA
CTTGAACATGCCTTCAGCAAGGCTCAAACTTATTTGGGAATTGAAAAGCTGTTAGATCCTGAAGATGTTG
CCGTTCAGCTTCCTGACAAGAAATCCATAATTATGTATTTAACATCTTTGTTTGAGGTGCTACCTCAGCA
AGTCACCATAGAC
>translated aa sequence
MAKYGEHEASPDNGQNEFSDIIKSRSDEHNDVQKKTFTKWINARFSKSGKPPINDMFTDL
KDGRKLLDLLEGLTGTSLPKERGSTRVHALNNVNRVLQVLHQNNVELVNIGGTDIVDGNH
KLTLGLLWSIILHWQVKDVMKDVMSDLQQTNSEKILLSWVRQTTRPYSQVNVLNFTTSWT
DGLAFNAVLHRHKPDLFSWDKVVKMSPIERLEHAFSKAQTYLGIEKLLDPEDVAVQLPDK
KSIIMYLTSLFEVLPQQVTIDCell Motil Cytoskeleton. 2007 Nov;64(11):822-32.
Versatile fluorescent probes for actin filaments based on the actin-binding domain of utrophin.
Burkel BM, von Dassow G, Bement WM.Department of Zoology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Actin filaments (F-actin) are protein polymers that undergo rapid assembly and disassembly and control an enormous variety of cellular processes ranging from force production to regulation of signal transduction. Consequently, imaging of F-actin has become an increasingly important goal for biologists seeking to understand how cells and tissues function. However, most of the available means for imaging F-actin in living cells suffer from one or more biological or experimental shortcomings. Here we describe fluorescent F-actin probes based on the calponin homology domain of utrophin (Utr-CH), which binds F-actin without stabilizing it in vitro. We show that these probes faithfully report the distribution of F-actin in living and fixed cells, distinguish between stable and dynamic F-actin, and have no obvious effects on processes that depend critically on the balance of actin assembly and disassembly. (c) 2007 Wiley-Liss, Inc.
Now that Bio Marin have stopped trials due to the plasma not reaching higher enough levels to increase the expression of utrophin will others still continue this research or try to find an alternative therapy.
Regards C.M. (future carer of DMD boy)