Method and kit for identifying a translation initiation site on an mrna
AssigneeCornell University
InventorsShu-Bing Qian, Sooncheol Lee, Botao Liu
Life of this patent
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Abstract
The present invention relates to a method and kit for identifying a translation initiation site on an mRNA. The method involves contacting a first mRNA with a first translation inhibitor to preferentially stabilize one or more initiation ribosomes at translation initiation sites on the first mRNA. A second mRNA is contacted with a second translation inhibitor different from the first translation inhibitor to stabilize one or more initiation ribosomes and one or more elongation ribosomes on the second mRNA. The location of ribosomes stabilized on the first mRNA is compared to the location of ribosomes stabilized on the second mRNA.
Claims27
A method for identifying a translation initiation site on an mRNA, said method comprising:
- providing a first mRNA in an environment suitable for translation;
- contacting the first mRNA with a first translation inhibitor to preferentially stabilize one or more initiation ribosomes at translation initiation sites on the first mRNA;
- providing a second mRNA in an environment suitable for translation, wherein the second mRNA has a nucleotide sequence that is substantially similar to a nucleotide sequence of the first mRNA;
- contacting the second mRNA with a second translation inhibitor different from the first translation inhibitor to stabilize one or more initiation ribosomes and one or more elongation ribosomes on the second mRNA;
- and
- comparing the location of ribosomes stabilized on the first mRNA to the location of ribosomes stabilized on the second mRNA, wherein ribosomes stabilized at a location on the first mRNA at a higher density than ribosomes stabilized at the same location on the second mRNA identifies the location as a translation initiation site on the first and second mRNAs.
A kit for identifying a translation initiation site on an mRNA, said kit comprising:
- a first translation inhibitor capable of preferentially stabilizing initiation ribosomes at translation initiation sites on an mRNA;
- a second translation inhibitor different from the first translation inhibitor, wherein the second translation inhibitor is capable of stabilizing initiation ribosomes and elongation ribosomes on an mRNA;
- and
- instructions for (i) contacting a first mRNA with the first translation inhibitor and a second mRNA with the second translation inhibitor and (ii) comparing the location of ribosomes stabilized on the first mRNA to ribosomes stabilized on the second mRNA to identify translation initiation sites on the first and second mRNAs.
Dependencies are inferred from the claim text.