Methods for mapping personalized translatome
AssigneeNorthwestern University
InventorsQianru LI, Haiwang YANG, Zhe JI
Life of this patent
Estimating the term…
Abstract
A low-input RNase footprinting approach is described for the rapid quantification of ribosome-protected fragments with 1,000-100,000 cells. The assay uses a simplified procedure for capturing ribosome-RNA complexes based on optimized RNase digestion. It simultaneously maps cytosolic and mitochondrial translation with single-nucleotide resolution.
Claims23
A method of RNAse footprinting comprising contacting a biological sample comprising cells with an RNAse in an amount sufficient to protect translational ribosome-bound fragments, sequencing the translational ribosome-bound fragments, and mapping the ribosome profile for each of the translational ribosome-bound fragments, wherein the number of cells in the biological sample is less than about 100,000, and wherein said method does not comprise ribosome isolation by centrifugation or immunoprecipitation.
A method of preparing open reading frame-coding RNA fragments from a biological sample comprising contacting the biological sample with an RNAse in an amount sufficient to protect translational ribosome-bound fragments from digestion by the RNAse, isolating the translational ribosome-bound fragments without centrifugation, and sequencing the fragments coding an open reading frame.
-32. (canceled)
A method of low input RNase footprinting comprising contacting a biological sample comprising 50,000 or less cells with an RNase in an amount sufficient to protect translational ribosome-bound fragments, sequencing the translational ribosome-bound fragments, and mapping the ribosome profile for each of the translational ribosome-bound fragments, wherein said method does not comprise centrifugation.
Dependencies are inferred from the claim text.