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US 10,501,779 B2Granted
Oligonucleotide trapping
AssigneePresident and Fellows of Harvard College
InventorsChao-ting Wu, Brian Beliveau
Life of this patent
Estimating the term…
Abstract
Novel methods and compositions for identifying one or more factors associated with a nucleic acid sequence (e.g., DNA and/or RNA) of interest are provided.
Claims26
A method for identifying one or more factors bound to one or more targeting moieties of a plurality of oligonucleotide paints in a biological sample by in situ hybridization, comprising the steps of:
- hybridizing the plurality of oligonucleotide paints to different chromosomal regions of a chromosome in the biological sample by in situ hybridization, thereby forming hybridized oligonucleotide paints, wherein each of the oligonucleotide paints comprises a 5′ primer sequence, a nucleotide sequence capable of hybridizing to a specific region of the chromosome, 3′ primer sequence, and one or more targeting moieties, and the nucleotide sequence capable of hybridizing to the specific region of the chromosome is located between the 5′ primer sequence and the 3′ primer sequence and is different in each of the oligonucleotide paints and the 5′ primer sequence and the 3′ primer sequence comprises either nucleotide sequences of a universal primer or nucleotide sequences fully complementary to the universal primer;
- removing the oligonucleotide paints unbound to the different chromosomal regions of the chromosome from the hybridized oligonucleotide paints;
- covalently attaching the targeting moieties of the hybridized oligonucleotide paints to the one or more factors in the biological sample;
- and
- identifying the one or more factors bound to the one or more targeting moieties of the plurality of oligonucleotide paints in the biological sample by identifying the factors attached to the targeting moieties of the hybridized oligonucleotide paints.
2 more elements
A method for identifying one or more factors bound to one or more targeting moieties of a plurality of oligonucleotide paints in a biological sample by in situ hybridization, comprising the steps of:
- hybridizing the plurality of oligonucleotide paints to different regions of genomic DNA or mitochondrial DNA in the biological sample by in situ hybridization, thereby forming hybridized oligonucleotide paints, wherein each of the oligonucleotide paints comprises a 5′ primer sequence, a nucleotide sequence capable of hybridizing to a specific region of the genomic DNA or the mitochondrial DNA, 3′ primer sequence, and one or more targeting moieties, and the nucleotide sequence capable of hybridizing to the specific region of the genomic DNA or the mitochondrial DNA is located between the 5′ primer sequence and the 3′ primer sequence and is different in each of the oligonucleotide paints and the 5′ primer sequence and the 3′ primer sequence comprises either nucleotide sequences of a universal primer or nucleotide sequences fully complementary to the universal primer;
- removing the oligonucleotide paints unbound to the different regions of the genomic DNA or the mitochondrial DNA from the hybridized oligonucleotide paints;
- covalently attaching the targeting moieties of the hybridized oligonucleotide paints to the one or more factors in the biological sample;
- and
- identifying the one or more factors bound to the one or more targeting moieties of the plurality of oligonucleotide paints in the biological sample by identifying the factors attached to the targeting moieties of the hybridized oligonucleotide paints.
2 more elements
A method for identifying one or more factors bound to one or more targeting moieties of a plurality of oligonucleotide paints in a biological sample by in situ hybridization, comprising the steps of:
- hybridizing the plurality of oligonucleotide paints to different regions of genomic DNA or mitochondrial DNA in the biological sample by in situ hybridization, thereby forming hybridized oligonucleotide paints, wherein the plurality of oligonucleotide paints has retrievable moieties, and wherein each of the oligonucleotide paints comprises a 5′ primer sequence, a nucleotide sequence capable of hybridizing to a specific region of the genomic DNA or the mitochondrial DNA, 3′ primer sequence, and one or more targeting moieties, and the nucleotide sequence capable of hybridizing to the specific region of the genomic DNA or the mitochondrial DNA is located between the 5′ primer sequence and the 3′ primer sequence and is different in each of the oligonucleotide paints and the 5′ primer sequence and the 3′ primer sequence comprises either nucleotide sequences of a universal primer or nucleotide sequences fully complementary to the universal primer;
- removing the oligonucleotide paints unbound to the different regions of the genomic DNA or mitochondrial DNA from the hybridized oligonucleotide paints;
- covalently attaching the targeting moieties of the hybridized oligonucleotide paints to the one or more factors in the biological sample;
- and
- identifying the one or more factors bound to the one or more targeting moieties of the plurality of oligonucleotide paints in the biological sample by identifying the factors attached to the targeting moieties of the hybridized oligonucleotide paints.
2 more elements
Dependencies are inferred from the claim text.