Extbase Variable Dump
array(14 items)
   uid => 4 (integer)
   title => 'Led-Seq: ligation-enhanced double-end sequence-based structure analysis of 
      RNA
' (79 chars) abstract => 'Structural analysis of RNA is an important and versatile tool to investigate
       the function of this type of molecules in the cell as well as in vitro. Sev
      eral robust and reliable procedures are available, relying on chemical modif
      ication inducing RT stops or nucleotide misincorporations during reverse tra
      nscription. Others are based on cleavage reactions and RT stop signals. Howe
      ver, these methods address only one side of the RT stop or misincorporation
      position. Here, we describe Led-Seq, a new approach based on lead-induced cl
      eavage of unpaired RNA positions, where both resulting cleavage products are
      
      
      A ligases. In a deep sequencing analysis, the cleavage sites are identified
      as ligation positions, avoiding possible false positive signals based on pre
      mature RT stops. With a benchmark set of transcripts in Escherichia coli, we
       show that Led-Seq is an improved and reliable approach based on metal ion-i
      nduced phosphodiester hydrolysis to investigate RNA structures in vivo.
' (1135 chars) authors => array(7 items) 0 => array(3 items) last_name => 'Kolberg' (7 chars) first_name => 'Tim' (3 chars) sorting => 1 (integer) 1 => array(3 items) last_name => 'Von Lohneysen' (13 chars) first_name => 'Sarah' (5 chars) sorting => 2 (integer) 2 => array(3 items) last_name => 'Ozerova' (7 chars) first_name => 'Iuliia' (6 chars) sorting => 3 (integer) 3 => array(3 items) last_name => 'Wellner' (7 chars) first_name => 'Karolin' (7 chars) sorting => 4 (integer) 4 => array(3 items) last_name => 'Hartmann' (8 chars) first_name => 'Roland K.' (9 chars) sorting => 5 (integer) 5 => array(3 items) last_name => 'Stadler' (7 chars) first_name => 'Peter Florian' (13 chars) sorting => 6 (integer) 6 => array(3 items) last_name => 'Mörl' (5 chars) first_name => 'Mario' (5 chars) sorting => 7 (integer) type => '0' (1 chars) keywords => 'Genetics' (8 chars) year => 2023 (integer) affiliation => 0 (integer) link_paper => 'https://academic.oup.com/nar/article-pdf/51/11/e63/50669469/gkad312.pdf' (71 chars) link_supplements => '' (0 chars) file_published => 0 (integer) journal => 'Nucleic Acids Research' (22 chars) doi => '10.1093/nar/gkad312' (19 chars) preprint => '-1' (2 chars)

Led-Seq: ligation-enhanced double-end sequence-based structure analysis of RNA

2023: Tim Kolberg; Sarah Von Lohneysen; Iuliia Ozerova; Karolin Wellner; Roland K. Hartmann; Peter Florian Stadler; Mario Mörl
In: Nucleic Acids Research
Link to Publication DOI: 10.1093/nar/gkad312

Structural analysis of RNA is an important and versatile tool to investigate the function of this type of molecules in the cell as well as in vitro. Several robust and reliable procedures are available, relying on chemical modification inducing RT stops or nucleotide misincorporations during reverse transcription. Others are based on cleavage reactions and RT stop signals. However, these methods address only one side of the RT stop or misincorporation position. Here, we describe Led-Seq, a new approach based on lead-induced cleavage of unpaired RNA positions, where both resulting cleavage products are investigated. The RNA fragments carrying 2′, 3′-cyclic phosphate or 5′-OH ends are selectively ligated to oligonucleotide adapters by specific RNA ligases. In a deep sequencing analysis, the cleavage sites are identified as ligation positions, avoiding possible false positive signals based on premature RT stops. With a benchmark set of transcripts in Escherichia coli, we show that Led-Seq is an improved and reliable approach based on metal ion-induced phosphodiester hydrolysis to investigate RNA structures in vivo.

Keywords: Genetics