FLAM-Seq: Genome-wide full-length mRNA and poly(A)-tail sequencing
The information about mRNA isoform expression including respective poly(A)-tail sequence (actual sequence length and sequence composition) is invaluable for the in-depth understanding of cellular gene regulation and RNA biology. Currently, different assay systems and technologies attain only long-read sequences or poly (A)-length estimation in a separate manner. Hence, it remains difficult to acquire this comprehensive information of the entire “mRNA architecture” (assembled coding regions, non-coding regions and corresponding poly(A)-tail) at once for a given gene on a quantitative level in a genome-wide transcriptomics approach. Even long-read cDNA sequencing technologies are limited in their capability to obtain the full sequencing information.
FLAM-Seq: Example of long reads including poly-A-Tail (in blue)
The invention provides an innovative solution for cDNA library preparation, enabling transcriptome-wide sequencing of full-length mRNAs including respective poly(A)-tail. FLAM-Seq (Full-length poly(A) mRNA sequencing) comprises a versatile protocol, enabling the annotation of mRNA isoforms and their abundance in conjunction with respective poly(A)-tail length. Instead of poly(A)-length estimation, FLAM-Seq reveals the global distribution of poly(A)-length and ultimately uncovers alternative nucleotide compositions within the poly(A)-sequence. No sequencing bias occurs when determining long Adenosine-sequence stretches. Sequencing of the “Full-length”-library based on this method guarantees high reproducibility with long comprehensive reads.
This opportunity is available for co-development or in-licensing.
Protocols for the methodology have been established, optimized and validated.
European Patent application EP18198248 has been filed with priority of October 2018.
Manuscript in preparation.