TATL-Seq
Translation-Associated Transcript Leader Sequencing
TATL-Seq works in conjunction with TL-Seq to target the sequence around the 5′-UTRs of mRNA attached to polysomes (Arribere et a., 2013). RNA from polysome gradient fractions is extracted, poly(A)-selected, and fragmented. Next, the TL-Seq protocol is followed to isolate and sequence the 5′-UTRs.
Advantages:
- Sequences 5′-UTRs and identifies variants
- TATL-Seq enables de novo transcript leader annotation while simultaneously testing their translational activity in a single experiment
Disadvantages:
- Selects short nucleotide fragments (50_80 nt)
- Labor-intensive; requires large amounts of starting material
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Bangru S. and Kalsotra A. Advances in analyzing RNA diversity in eukaryotic transcriptomes: peering through the Omics lens. F1000Research. 2016;5:2668
Smith J. E. and Baker K. E. Nonsense-mediated RNA decay–a switch and dial for regulating gene expression. Bioessays. 2015;37:612-623
References:
Arribere J. A. and Gilbert W. V. Roles for transcript leaders in translation and mRNA decay revealed by transcript leader sequencing. Genome Res. 2013;23:977-987