Nucleic acid-controlled catalytic rnas for trigger-responsive regulation
US-2024425855-A1 · Dec 26, 2024 · US
Methods of making guide RNA
US11261439B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11261439-B2 |
| Application number | US-201615760372-A |
| Country | US |
| Kind code | B2 |
| Filing date | Sep 16, 2016 |
| Priority date | Sep 18, 2015 |
| Publication date | Mar 1, 2022 |
| Grant date | Mar 1, 2022 |
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Abstract
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A nucleic acid construct is provided that encodes two or more or a plurality of spacer sequences separated by restriction endonuclease recognition site. A plurality of such nucleic acid sequences are provided as a library for making guide RNAs for use with CRISPR/Cas systems.
First claim
Opening claim text (preview).
The invention claimed is: 1. A method of making a library of nucleic acids encoding guide RNA spacer sequences wherein the guide RNA spacer sequences are complementary to corresponding target nucleic acid sequences comprising synthesizing a plurality of nucleic acid sequences each encoding (1) a plurality of spacer sequences including a pair of end spacer sequences, (2) inner restriction endonuclease cut sites, wherein the restriction endonuclease cut sites separate each of the plurality of spacer sequences from each other, (3) a pair of outer restriction endonuclease cut sites flanking the pair of end spacer sequences, and (4) a pair of amplification primer binding sites flanking the outer restriction endonuclease cut sites, and wherein the outer restriction endonuclease cut sites are different from the inner restriction endonuclease cut sites. 2. The method of claim 1 wherein each nucleic acid sequence encodes 3 to 20 spacer sequences. 3. The method of claim 1 wherein the inner restriction endonuclease cut sites are different from each other. 4. The method of claim 1 wherein the plurality of nucleic acid sequences are synthesized on a support. 5. The method of claim 4 wherein the plurality of nucleic acid sequences are removed from the support.
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Classifications
Mutagenizing nucleic acids · CPC title
- C12N15/1093Primary
General methods of preparing gene libraries, not provided for in other subgroups · CPC title
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Frequently asked questions
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- What does patent US11261439B2 cover?
- A nucleic acid construct is provided that encodes two or more or a plurality of spacer sequences separated by restriction endonuclease recognition site. A plurality of such nucleic acid sequences are provided as a library for making guide RNAs for use with CRISPR/Cas systems.
- Who is the assignee on this patent?
- Harvard College
- What technology area does this patent fall under?
- Primary CPC classification C12N15/1093. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Mar 01 2022 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).