title: Applications of PLOR in labeling large RNAs at specific sites.
abstract: Incorporation of modified or labeled nucleotides at specific sites in RNAs is critical for gaining insights into the structure and function of RNAs. Preparation of site-specifically labeled large RNAs in amounts suitable for structural or functional studies is extremely difficult using current methodologies. The position-selective labeling of RNA, PLOR, is a recently developed method that makes such syntheses possible. PLOR allows incorporation of various probes, including (2)D/(13)C/(15)N-isotopic labels, Cy3/Cy5/Alexa488/Alexa555 fluorescent dyes, biotin and other chemical groups, into specific positions in long RNAs. Here, we describe in detail the use of PLOR to label RNAs at specific segment(s) or discrete sites.
authors.affiliation: Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA. Electronic address: liuy13@mail.nih.gov.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Structural Biophysics Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.affiliation: Structural Biophysics Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors.forename: Marzena
authors.initials: M
authors.lastname: Dyba
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center, San Antonio, TX 78229, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Sousa
authors.affiliation: Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Jason R
authors.initials: JR
authors.lastname: Stagno
authors.affiliation: Protein-Nucleic Acid Interaction Section, Structural Biophysics Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.
authors.forename: Yun-Xing
authors.initials: YX
authors.lastname: Wang
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry; D055029:Inverted Repeat Sequences; D012313:RNA / Q000737:chemistry*; D013194:Staining and Labeling; D014764:Viral Proteins / Q000737:chemistry
chemical_list: C417561:Alexa 488 hydrazide; D005456:Fluorescent Dyes; D006834:Hydrazines; D014764:Viral Proteins; D012313:RNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases
keywords: PLOR; RNA; Specific labeling