Carrocci Tucker J, Lohe Lea, Ashton Matthew J, Höbartner Claudia, Hoskins Aaron A
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Dr., Madison, WI 53706, USA.
Institute for Organic and Biomolecular Chemistry, Georg-August-University Göttingen, Tammannstr. 2, 37077 Göttingen, Germany.
Chem Commun (Camb). 2017 Nov 14;53(88):11992-11995. doi: 10.1039/c7cc06703h. Epub 2017 Oct 6.
The 10DM24 deoxyribozyme can site-specifically label RNAs with fluorophore-GTP conjugates; however, the 2',5'-branched RNA linkage is readily cleaved by debranchase. To prevent loss of labels upon cleavage, we synthesized phosphorothioate-modified, fluorescent GTP derivatives and elaborated conditions for their incorporation by 10DM24. RNAs labeled with fluorescent derivatives of Sp-GTP were found to be resistant to debranchase.
10DM24脱氧核酶可以用荧光团-GTP偶联物对RNA进行位点特异性标记；然而，2'，5'-分支的RNA连接很容易被去分支酶切割。为了防止切割时标记丢失，我们合成了硫代磷酸酯修饰的荧光GTP衍生物，并详细阐述了10DM24将其掺入的条件。发现用Sp-GTP荧光衍生物标记的RNA对去分支酶具有抗性。
