Open AccessLow-bias ncRNA libraries using ordered two-template relay: Serial template jumping by a modified retroelement reverse transcriptase
Author(s) -
Heather Upton,
Lucas Ferguson,
Morayma M. Temoche-Diaz,
Xiao Man Liu,
Sydney C. Pimentel,
Nicholas T. Ingolia,
Randy Schekman,
Kathleen Collins
Publication year - 2021
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.2107900118
Subject(s) - biology , reverse transcriptase , complementary dna , computational biology , rna , long terminal repeat , primer (cosmetics) , retrotransposon , genetics , gene , gene expression , transposable element , chemistry , organic chemistry , genome
Significance Retrotransposons are noninfectious, mobile genetic elements that proliferate in host genomes via an RNA intermediate that is copied into DNA by a reverse transcriptase (RT) enzyme. RTs are important for biotechnological applications involving information capture from RNA since RNA is first converted into complementary DNA for detection or sequencing. Here, we biochemically characterized RTs from two retroelements and uncovered several activities that allowed us to design a streamlined, efficient workflow for determining the inventory of RNA sequences in processed RNA pools. The unique properties of nonretroviral RT activities obviate many technical issues associated with current methods of RNA sequence analysis, with wide applications in research, biotechnology, and diagnostics.