Gene silencing
AssigneePlant Bioscience Limited
InventorsDavid Charles Baulcombe, Andrew John Hamilton
Life of this patent
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Abstract
Methods are disclosed for screening for the occurrence of gene silencing (e.g., post transcriptional gene silencing) in an organism. Also provided are methods for isolating silencing agents so identified.
Claims16
A method of detecting the silencing of a target gene in a plant, wherein said silencing is initiated by introduction of an exogenous nucleic acid, which method comprises the steps of:
- (i) obtaining a sample of material from said plant,
- (ii) producing a nucleic acid extract from said sample,
- (iii) analyzing said extract such as to determine the presence or absence of short RNA molecules which are 21-25 nucleotides in length (SRMs) in said extract,
- (iv) characterizing any SRMs which are present in said extract such as to determine sequence identity or similarity with said target gene, and
- (v) correlating the presence of said SRMs having sequence identity or similarity with said target gene in the extract with the occurrence of gene silencing in said plant.
A process for isolating one or more RNA molecules associated with target gene silencing from a sample of material from a plant, wherein the RNA molecules are SRMs which share sequence identity with the target gene, and wherein said silencing is initiated by introduction of an exogenous nucleic acid, which process comprises the steps of:
- (a) producing a nucleic acid extract from said sample,
- (b) purifying said extract to produce purified RNA molecules which are 21-25 nucleotides in length by carrying out at least one purification step selected from the following steps (i) filtration;
- (ii) differential precipitation (iii) ion exchange chromatography, such as to isolate said SRMs.
Dependencies are inferred from the claim text.