High-throughput translational profiling with riboPLATE-seq.

Autor: Metz JB; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, 10032, USA.; Medical Scientist Training Program, Columbia University Irving Medical Center, New York, NY, 10032, USA., Hornstein NJ; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, 10032, USA.; Medical Scientist Training Program, Columbia University Irving Medical Center, New York, NY, 10032, USA.; MD Anderson Cancer Center, Houston, TX, 77030, USA., Sharma SD; Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, NY, 10065, USA., Worley J; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, 10032, USA., Gonzalez C; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, 10032, USA., Sims PA; Department of Systems Biology, Columbia University Irving Medical Center, New York, NY, 10032, USA. pas2182@columbia.edu.; Department of Biochemistry and Molecular Biophysics, Columbia University Irving Medical Center, New York, NY, 10032, USA. pas2182@columbia.edu.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2022 Apr 05; Vol. 12 (1), pp. 5718. Date of Electronic Publication: 2022 Apr 05.
DOI: 10.1038/s41598-022-09638-3
Abstrakt: Protein synthesis is dysregulated in many diseases, but we lack a systems-level picture of how signaling molecules and RNA binding proteins interact with the translational machinery, largely due to technological limitations. Here we present riboPLATE-seq, a scalable method for generating paired libraries of ribosome-associated and total mRNA. As an extension of the PLATE-seq protocol, riboPLATE-seq utilizes barcoded primers for pooled library preparation, but additionally leverages anti-rRNA ribosome immunoprecipitation on whole polysomes to measure ribosome association (RA). We compare RA to its analogue in ribosome profiling and RNA sequencing, translation efficiency, and demonstrate both the performance of riboPLATE-seq and its utility in detecting translational alterations induced by specific inhibitors of protein kinases.
(© 2022. The Author(s).)
Databáze: MEDLINE
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