Compositions and methods for immunooncology
US-2024417722-A1 · Dec 19, 2024 · US
CRISPR oligonucleotides and gene editing
US9879283B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9879283-B2 |
| Application number | US-201514879872-A |
| Country | US |
| Kind code | B2 |
| Filing date | Oct 9, 2015 |
| Priority date | Oct 9, 2014 |
| Publication date | Jan 30, 2018 |
| Grant date | Jan 30, 2018 |
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- Title
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- Abstract
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Abstract
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The present disclosure generally relates to compositions and methods for the genetic modification of cells. In particular, the disclosure relates to CRISPR reagents and the use of such reagents.
First claim
Opening claim text (preview).
What is claimed is: 1. A method for producing a nucleic acid molecule, the method comprising performing polymerase chain reaction (PCR) in a reaction mixture comprising: (i) a double stranded nucleic acid segment comprising a first terminus and a second terminus, (ii) a first oligonucleotide comprising a first terminus and a second terminus, wherein the second terminus of the first oligonucleotide is capable of hybridizing to the first terminus of the double stranded nucleic acid segment, and (iii) a second oligonucleotide comprising a first terminus and a second terminus, wherein the second terminus of the second oligonucleotide is capable of hybridizing to the first terminus of the first oligonucleotide, to produce the nucleic acid molecule, wherein the product nucleic acid molecule contains a promoter suitable for in vitro transcription at or near one terminus and encodes a CRISPR RNA, and wherein the reaction mixture further comprises a first primer and a second primer, wherein the first primer is capable of hybridizing at or near the first terminus of the second oligonucleotide and the second primer is capable of hybridizing at or near the second terminus of the double stranded nucleic acid segment. 2. The method of claim 1 , wherein the CRISPR RNA is a guide RNA. 3. The method of claim 2 , wherein the nucleic acid molecule produced by the PCR reaction is from 70 to 150 base pairs in length. 4. The method of claim 1 , wherein the CRISPR RNA is from 35 to 150 nucleotides in length. 5. The method of claim 1 , wherein the CRISPR RNA has at least two hairpin turns. 6. The method of claim 1 , wherein the CRISPR RNA is a crRNA. 7. The method of claim 1 , wherein the CRISPR RNA is a tracrRNA. 8. The method of claim 1 , wherein the promoter is a T7, T3 or SP6 promoter. 9. The method of claim 1 , wherein the first oligonucleotide or the second oligonucleotide is between 35 and 40 nucleotides in length.
Assignees
Inventors
Classifications
incorporating a promoter sequence · CPC title
Fusion with another nucleic acid · CPC title
- C12N15/907Primary
in mammalian cells · CPC title
Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title
Mutagenizing nucleic acids · CPC title
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Frequently asked questions
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- What does patent US9879283B2 cover?
- The present disclosure generally relates to compositions and methods for the genetic modification of cells. In particular, the disclosure relates to CRISPR reagents and the use of such reagents.
- Who is the assignee on this patent?
- Life Technologies Corp, Thermo Fisher Scient Geneart Gmbh
- What technology area does this patent fall under?
- Primary CPC classification C12N15/907. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Jan 30 2018 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).