Method for Sensitive Detection of Target DNA Using Target-Specific Nuclease
First Claim
1. A method for analyzing a genotype comprising:
- (i) removing particular genotype DNA in an isolated sample, in which two or more genotypes of DNA are mixed, by cleaving the particular genotype DNA using a nuclease specific to the particular genotype DNA; and
(ii) analyzing other genotype DNA present in the isolated sample, in which the particular genotype DNA has been removed.
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Abstract
The present invention relates to a method for analyzing a genotype using a target-specific nuclease and, specifically, to a method for diagnosing cancer or analyzing a genotype by removing wild type DNA or particular genotype DNA using a target-specific nuclease or a variant thereof to amplify or concentrate only a small amount of DNA which has a difference in variation, such as a mutation, or genotype, and to a method for separating target DNA sesusing a target-specific nuclease or a variant thereof. Such methods are novel paradigm methods contrary to existing simple target-specific nucleases for post-PCR recognition of normal genotype and carcinogenic genotype, and can be favorably used in the early diagnosis of cancer or analysis of similar genotypes.
158 Citations
26 Claims
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1. A method for analyzing a genotype comprising:
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(i) removing particular genotype DNA in an isolated sample, in which two or more genotypes of DNA are mixed, by cleaving the particular genotype DNA using a nuclease specific to the particular genotype DNA; and (ii) analyzing other genotype DNA present in the isolated sample, in which the particular genotype DNA has been removed. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method for analyzing a genotype comprising:
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(i) removing other genotype DNA by cleaving the other genotype DNA while masking particular genotype DNA in an isolated sample, in which two or more genotypes of DNA are mixed, for protection from cleavage by RGEN capable of recognizing the other genotype DNA, using a guide RNA (gRNA) specific to the particular genotype DNA and an inactivated Cas nuclease protein (dCas); and (ii) analyzing the particular genotype DNA present in the sample, in which the other genotype DNA has been removed. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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22. A method for analyzing a genotype of DNA in an isolated sample comprising:
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(i) removing DNA of a non-pathogenic bacterium or virus in the isolated sample by cleaving the DNA of the non-pathogenic bacterium or virus by treating the DNA of the non-pathogenic bacterium or virus with a nuclease specific to the DNA of the non-pathogenic bacterium or virus, in the isolated sample having DNA of bacteria or viruses; and (ii) analyzing DNA of a pathogenic bacterium or virus in the isolated sample in which the DNA of the non-pathogenic bacterium or virus has been removed. - View Dependent Claims (23, 24)
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25. A method for analyzing a genotype of DNA in an isolated sample comprising:
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(i) removing DNA of a non-pathogenic bacterium or virus in the isolated sample by cleaving the DNA of the non-pathogenic bacterium or virus while masking DNA of a pathogenic bacterium or virus for protection from cleavage by RGEN specific to the non-pathogenic bacterium or virus, using a guide RNA specific to the pathogenic bacterium or virus and an inactivated Cas nuclease protein (dCas); and (ii) analyzing the DNA of the pathogenic bacterium or virus in the isolated sample in which the DNA of the non-pathogenic bacterium or virus has been removed. - View Dependent Claims (26)
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Specification