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FLAM-Seq: Genome-wide full-length mRNA and poly(A)-tail sequencing

Reference Number TO 03-00470

Challenge

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)
FLAM-Seq: Example of long reads including poly-A-Tail (in blue)

Technology

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.

Commercial Opportunity

This opportunity is available for co-development or in-licensing.

Developmental Status

Protocols for the methodology have been established, optimized and validated.

Patent Situation

European Patent application EP18198248 has been filed with priority of October 2018. 

Further Reading

Manuscript in preparation.