Studying Selenoprotein mRNA Translation Using RNA-Seq and Ribosome Profiling.
Methods Mol Biol
; 1661: 103-123, 2018.
Article
in En
| MEDLINE
| ID: mdl-28917040
ABSTRACT
Deep sequencing of ribosome protected mRNA footprints, also called ribosome profiling or Ribo-Seq, is a relatively new methodology well suited to address questions regarding the mechanisms and efficiency of protein expression. Specifically, the ability of this technique to quantify ribosome abundance with codon resolution enables experiments aimed at studying many aspects of translation, including gene-specific translational efficiency, translation of regulatory upstream short open reading frames, sites of ribosome pausing, and most importantly for selenoproteins, the efficiency by which UGA codons are redefined to encode selenocysteine. Here, we describe a streamlined protocol that was developed in our lab to process mammalian tissue to produce the requisite matched ribosome profiling and RNA-Seq libraries for deep sequencing.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Ribosomes
/
Protein Biosynthesis
/
Selenoproteins
/
High-Throughput Nucleotide Sequencing
Limits:
Animals
Language:
En
Journal:
Methods Mol Biol
Year:
2018
Document type:
Article