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Title:
Efficient 5' Cap-Dependent RNA Purification: Use in Identifying and Studying Subsets of RNA | SpringerLink
Description:
Microarray-based screening technologies have revealed a larger than expected diversity of gene expression profiles for many cells, tissues, and organisms. The complexity of RNA species, defined by their molecular structure, represents a major new development in...
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Topics {✒️}

post-transcriptional gene regulation cap-binding protein eif4e post-transcriptional regulation springer science+business media micro-rna genes mir15 gene expression profiles full-length cdna ends privacy choices/manage cookies karolinska university press m7g cap analogues antisense rna transcripts differential selection approaches gene expression studies + rna-interacting proteins increased binding affinity functional eif-4ehuman colorado state university disclosing hidden transcripts high-affinity variant author information authors journal finder publish rna polymerase ii high-resolution map european economic area protein synthesis artificial tethered enzyme chronic lymphocytic leukemia s4-h2 loop spivak-kroizman sepharose-coupled m7gdp nih ro1 ca063640 humana press carries genetic information mammalian exosome mediates conditions privacy policy protocol cite protocol bajak laminar shear stress nucleic acids res international university line kansas medical center usa edyta general translational repression accepting optional cookies detection methods au-rich elements tetrahymena thermophila mrnas cytoplasmic processing bodies main content log 3’ inverse race-pcr

Schema {🗺️}

ScholarlyArticle:
      headline:Efficient 5' Cap-Dependent RNA Purification: Use in Identifying and Studying Subsets of RNA
      pageEnd:160
      pageStart:147
      image:https://media.springernature.com/w153/springer-static/cover/book/978-1-59745-033-1.jpg
      genre:
         Springer Protocols
      isPartOf:
         name:Post-Transcriptional Gene Regulation
         isbn:
            978-1-59745-033-1
            978-1-58829-783-9
         type:Book
      publisher:
         name:Humana Press
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:Edyta Z. Bajak
            affiliation:
                  name:University of Kansas Medical Center
                  address:
                     name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Curt H. Hagedorn
            affiliation:
                  name:University of Kansas Medical Center
                  address:
                     name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
                     type:PostalAddress
                  type:Organization
            type:Person
      keywords:RNA purification, 5′ RNA cap, m7G cap, eIF4E, microarray, mRNA, non-coding RNA, 3′ poly(A), deadenylation, cytoplasmic processing bodies
      description:Microarray-based screening technologies have revealed a larger than expected diversity of gene expression profiles for many cells, tissues, and organisms. The complexity of RNA species, defined by their molecular structure, represents a major new development in biology. RNA not only carries genetic information in the form of templates and components of the translational machinery for protein synthesis but also directly regulates gene expression as exemplified by micro-RNAs (miRNAs). Recent evidence has demonstrated that 5′ capped and 3′ polyadenylated ends are not restricted to mRNAs, but that they are also present in precursors of both miRNAs and some antisense RNA transcripts. In addition, as many as 40% of transcribed RNAs may lack 3′ poly(A) ends. In concert with the presence of a 5′ cap (m7 GpppN), the length of the 3′ poly(A) end plays a critical role in determining the translational efficiency, stability, and the cellular distribution of a specific mRNA. RNAs with short or lacking 3′ poly(A) ends, that escape isolation and amplification with oligo(dT)-based methods, provide a challenge in RNA biology and gene expression studies. To circumvent the limitations of 3′ poly(A)-dependent RNA isolation methods, we developed an efficient RNA purification system that binds the 5′ cap of RNA with a high-affinity variant of the cap-binding protein eIF4E. This system can be used in differential selection approaches to isolate subsets of RNAs, including those with short 3′ poly(A) ends that are likely targets of post-transcriptional regulation of gene expression. The length of the 3′ poly(A) ends can be defined using a rapid polymerase chain reaction (PCR)- based approach.
      datePublished:2008
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Book:
      name:Post-Transcriptional Gene Regulation
      isbn:
         978-1-59745-033-1
         978-1-58829-783-9
Organization:
      name:Humana Press
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         url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
         type:ImageObject
      name:University of Kansas Medical Center
      address:
         name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
         type:PostalAddress
      name:University of Kansas Medical Center
      address:
         name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
         type:PostalAddress
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      url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
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      name:Edyta Z. Bajak
      affiliation:
            name:University of Kansas Medical Center
            address:
               name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
               type:PostalAddress
            type:Organization
      name:Curt H. Hagedorn
      affiliation:
            name:University of Kansas Medical Center
            address:
               name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
               type:PostalAddress
            type:Organization
PostalAddress:
      name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
      name:Department of Medicine and Pharmacology, University of Kansas Medical Center, Kansas City, USA
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