Methods of depleting a target molecule from an initial collection of nucleic acids, and compositions and kits for practicing the same
Provided are methods of depleting a target nucleic acid from an initial collection of nucleic acids. Aspects of the methods include contacting the initial collection with a nucleic acid guided nuclease specific for the target nucleic acid in a manner sufficient to deplete the target nucleic acid from the initial collection. Depending on a given application, depletion of a target nucleic acid may vary, e.g., where depleting may include cleaving a target nucleic acid in, or selectively separating a target nucleic acid from, the initial collection of nucleic acids. Also provided are compositions and kits for practicing embodiments of the methods.
1. A method of selectively depleting target cDNAs from an RNA sequencing library sample, the method comprising:
obtaining an RNA sequencing library sample comprising both target cDNAs and non-target cDNAs, wherein the target cDNAs comprise cDNAs transcribed from ribosomal RNAs,
contacting the sample with a nucleic acid guided nuclease and a guide nucleic acid that guides the nucleic acid guided nuclease to the target cDNAs, wherein the nucleic acid guided nuclease, the guide nucleic acid, and the target cDNAs form a complex in which the guide nucleic acid specifically hybridizes with the target cDNAs, and
removing the target cDNAs from the sample by cleaving the target cDNAs with the nucleic acid guided nuclease in the complex, thereby selectively depleting the target cDNAs from the sample.
2. The method according to claim 1 , wherein the target cDNAs are double-stranded nucleic acids.
3. The method according to claim 1 , wherein the target cDNAs are single-stranded nucleic acids.
4. The method according to any of claim 1 , wherein the guide nucleic acid is RNA.
5. The method according to claim 4 , wherein the nucleic acid guided nuclease is a Cas nuclease.
6. The method of claim 4 , wherein the RNA is single-guide RNA specifically hybridized to human 5S, 5.8S, 18S, or 28S rRNAs.
7. The method according to claim 1 , wherein the guide nucleic acid is DNA.
8. The method according to claim 7 , wherein the nucleic acid guided nuclease is a Tth Ago nuclease.
9. The method according to claim 1 , wherein the method comprises further contacting the sample with one or more additional distinct nucleic acid guided nuclease.
10. The method of claim 9 , wherein the one or more additional distinct nucleic acid guide nucleases are guided by one or more distinct guide nucleic acids.
11. The method according to claim 1 , further comprising a step of squencing the sample.