An RNA-ligation-based RACE-PAT assay to monitor poly(A) tail length of mRNAs of interest

Fabienne Mauxion (1, 2) , Bertrand Séraphin (1, 2)

1 IGBMC - Institut de génétique et biologie moléculaire et cellulaire
2 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire

In eukaryotes, a non-templated polyadenosines (poly(A)) tail is added co-transcriptionally on almost every messenger RNA (mRNA). The length of this poly(A) tail changes during the lifetime of mRNAs and was shown in many circumstances to be an important factor controlling transcript fates. Yet, the measure of the length of this homogenous nucleotide sequence is technically challenging making it difficult to assess its dynamic variation. Here, we describe an RNA-ligation-based RACE-PAT (Rapid Amplification of cDNA End – Poly(A) Tail) assay to monitor poly(A) tail length of mRNAs of interest that we successfully used to study the function of the CCR4-NOT complex in mRNA deadenylation and its regulation by the antiproliferative BTG/Tob factors.

Domaines: Biochimie, Biologie Moléculaire

Fabienne Mauxion, Bertrand Séraphin. An RNA-ligation-based RACE-PAT assay to monitor poly(A) tail length of mRNAs of interest. Methods in Molecular Biology, pp.113-123, 2023, 978-1-0716-3481-3. ⟨10.1007/978-1-0716-3481-3_7⟩. ⟨hal-04259236⟩