Highly efficient in vitro translation of authentic affinity-purified messenger ribonucleoprotein complexes.

ABSTRACT: Cell-free systems are widely used to study mechanisms and regulation of translation, but the use of in vitro transcribed (IVT) mRNAs as translation substrates limits their efficiency and utility. Here, we present an approach for in vitro translation of messenger ribonucleoprotein (mRNP) complexes affinity purified in association with tagged mRNAs expressed in mammalian cells. We show that in vitro translation of purified mRNPs is much more efficient than that achieved using standard IVT mRNA substrates and is compatible with physiological ionic conditions. The high efficiency of affinity-purified mRNP in vitro translation is attributable to both copurified protein components and proper mRNA processing and modification. Further, we use translation inhibitors to show that translation of puri

SUBMITTER: Fritz SE 

PROVIDER: S-EPMC6004058 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature