title: Transglutaminase-mediated in situ hybridization (TransISH) system: a new methodology for simplified mRNA detection.
abstract: Detection and localization of specific DNA or RNA sequences in cells and tissues are of great importance for biological research, diagnosis, and environmental monitoring. However, the most common procedure for in situ hybridization employs laborious immunostaining techniques. In the present study, we report proof-of-concept for a new RNA-enzyme conjugated probe for the detection of mRNA on tissue sections with a simple procedure. An RNA probe modified with a specific dipeptide substrate of transglutaminase was prepared. Alkaline phosphatase was then covalently and site-specifically combined to the dipeptide-labeled RNA using microbial transglutaminase. The new RNA probe labeled with alkaline phosphatase was validated by in situ hybridization (ISH) and proved to be a sensitive and sequence specific probe for mRNA detection in tissues. The new transglutaminase-mediated ISH (TransISH) strategy is free from antigen-antibody reaction, leads to one-step signal amplification after hybridization, and thus will be widely applicable for highly sensitive nucleic acid detection.
authors: Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, Nishi-ku, Fukuoka, Japan.
authors: Momoko
authors: M
authors: Kitaoka
authors: Masayuki
authors: M
authors: Mitsumori
authors: Kounosuke
authors: K
authors: Hayashi
authors: Yoshiyuki
authors: Y
authors: Hiraishi
authors: Hisao
authors: H
authors: Yoshinaga
authors: Koji
authors: K
authors: Nakano
authors: Katsuyuki
authors: K
authors: Miyawaki
authors: Sumihare
authors: S
authors: Noji
authors: Masahiro
authors: M
authors: Goto
authors: Noriho
authors: N
authors: Kamiya
mesh_terms: D000818:Animals; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D011479:Protamines / Q000235:genetics; D020027:Pyrococcus furiosus / Q000201:enzymology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism*; D058949:Uromodulin / Q000235:genetics
chemical_list: D011479:Protamines; D012333:RNA, Messenger; D058949:Uromodulin; D011503:Transglutaminases