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US 2024/0254479 A1Published application
Engineered guide rna for optimized crispr/cas12f1 system and use thereof
AssigneeGenkore Inc.
InventorsYong-Sam KIM, Do Yon KIM
Life of this patent
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Abstract
In the present disclosure, there is provided an engineered Cas12f1 guide RNA for increasing intracellular gene editing activity of a CRISPR/Cas12f1 system by overcoming limitations of the prior art. The engineered Cas12f1 guide RNA is an engineered form of a naturally occurring guide RNA whose structure is partially modified. In addition, the engineered Cas12f1 guide RNA is characterized in that at least a portion of the scaffold region, which serves to interact with the Cas12f1 protein, is modified. The engineered scaffold region is characterized in that it is different from a scaffold region of a naturally occurring guide RNA.
Claims33
An engineered guide RNA for a CRISPR/Cas12f1 system, comprising:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUGAAAGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 7), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), and 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117);
- and
- 5′-AUGCAAC-3′.
46 more elements
An engineered guide RNA for a CRISPR/Cas12f1 system, comprising:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAAGA AAGUUGCAGAACCCGAAUAGACGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 315), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117), 5′-AACAAAUUCAUUUUGAAACGAAUGAAGGA-3′ (SEQ ID NO:
- 293), 5′-AACAAAUUCAUUUUUGAAAACGAAUGAAGGA-3′ (SEQ ID NO:
- 294), 5′-AACAAAUUCAUUUUUCGAAAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 295), 5′-AACAAAUUCAUUUUUCCGAAAAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 296), 5′-AACAAAUUCAUUUUUCCUGAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 297), 5′-AACAAAUUCAUUUUUCCUCGAAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 298), 5′-AACAAAUUCAUUUUUCCUCUGAAAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 299), 5′-AACAAAUUCAUUUUUCCUCUCGAAAGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 300), 5′-AACAAAUUCAUUUUUCCUCUCCGAAACGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 301), 5′-AACAAAUUCAUUUUUCCUCUCCAGAAACCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 302), 5′-AACAAAUUCAUUUUUCCUCUCCAAGAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 303), 5′-AACAAAUUCAUUUUUCCUCUCCAAUGAAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 304), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUGAAAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 305), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCGAAAGAACCCGAAUAGACGAAUGAAGG A-3′ (SEQ ID NO:
- 306), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGAAAAGAACCCGAAUAGACGAAUGAA GGA-3′ (SEQ ID NO:
- 307), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGGAAACAGAACCCGAAUAGACGAAUG AAGGA-3′ (SEQ ID NO:
- 308), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCGAAAGCAGAACCCGAAUAGACGAAU GAAGGA-3′ (SEQ ID NO:
- 309), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCAGAAAUGCAGAACCCGAAUAGACGA AUGAAGGA-3′ (SEQ ID NO:
- 310), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACGAAAUUGCAGAACCCGAAUAGAC GAAUGAAGGA-3′ (SEQ ID NO:
- 311), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAGAAAGUUGCAGAACCCGAAUAG ACGAAUGAAGGA-3′ (SEQ ID NO:
- 312), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAGAAAUUGCAGAACCCGAAUAGA CGAAUGAAGGA-3′ (SEQ ID NO:
- 313), and 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAAGAAAGUUGCAGAACCCGAAUA GACGAAUGAAGGA-3′ (SEQ ID NO:
- 314);
- and
- 5′-AUGCAAC-3′.
68 more elements
An engineered guide RNA for a CRISPR/Cas12f1 system, comprising:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer has a length of 10 to 50 nucleotides and has a sequence complementary to a target sequence, and
- a sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a first sequence represented by 5′-A-3′;
- a second sequence represented by 5′-CCGCUUCAC-3′ (SEQ ID NO:
- 432);
- a third sequence represented by 5′-UUAG-3′;
- a fourth sequence represented by 5′-AGUGAAGGUGG-3′ (SEQ ID NO:
- 433);
- a fifth sequence represented by 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sixth sequence represented by 5′-AACAAA-3′;
- a linker;
- a seventh sequence represented by 5′-GGA-3′;
- and
- an eighth sequence represented by 5′-AUGCAAC-3′.
15 more elements
An engineered guide RNA for a CRISPR/Cas12f1 system, comprising:
- an engineered scaffold region, and
- a spacer,
- wherein the spacer has a length of 10 to 50 nucleotides and has a sequence complementary to a target sequence, and
- a sequence of the engineered scaffold region comprises in a 5′ to 3′ direction:
- an engineered tracrRNA in which the following sequences are linked to each other:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9),
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- ID NO:
- 430)′ 5′-CCGCUUCACCUUAGGAGUGAAGGUG′-3′ (SEQ ID NO:
- 431)′ 5′-CCGCUUCACCAUUAGUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 38)′ 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 39)′ 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 40)′ 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 41)′ 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 42)′ 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 44)′ 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 45)′ 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 46)′ 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 47)′ 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUG′-3′(SEQ ID NO:
- 48)′ 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 49), an′ 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUG′-3′ (SEQ ID NO:
- 10),
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11), and
- a sequence selected from the group consisting of 5′-AACAAA-3′, 5′-AACAAAU-3′, 5′-AACAAAUU-3′, 5′-AACAAAUUC-3′, 5′-AACAAAUUCA-3′ (SEQ ID NO:
- 66), 5′-AACAAAUUCAU-3′ (SEQ ID NO:
- 67), 5′-AACAAAUUCAUU-3′ (SEQ ID NO:
- 68), and 5′-AACAAAUUCAUUU-3′ (SEQ ID NO:
- 12);
- and
- an engineered crRNA repeat sequence portion in which the following sequences are linked to each other:
- a sequence selected from the group consisting of 5′-GGA-3′, 5′-AGGA-3′, 5′-AAGGA-3′, 5′-GAAGGA-3′, 5′-UGAAGGA-3′, 5′-AUGAAGGA-3′, 5′-AAUGAAGGA-3′, and 5′-GAAUGAAGGA-3′ (SEQ ID NO:
- 14), and
- 5′-AUGCAAC-3′,
- wherein the 3′ end of the engineered crRNA repeat sequence is linked to the 5′ end of the spacer, and
- wherein one in which the sequence of the engineered tracrRNA is the same as 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUU-3′ (SEQ ID NO:
- 1) and the engineered crRNA repeat sequence is the same as 5′-GAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 3) is excluded.
48 more elements
An engineered CRISPR/Cas12f1 complex comprising:
- a Cas12f1 protein, and
- an engineered guide RNA,
- wherein the engineered guide RNA comprises:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUGAAAGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 7), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), and 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117);
- and
- 5′-AUGCAAC-3′.
49 more elements
A vector capable of expressing respective components of a CRISPR/Cas12f1 system, comprising:
- a first sequence comprising a nucleic acid sequence encoding a Cas12f1 protein;
- a first promoter sequence operably linked to the first sequence;
- a second sequence comprising a nucleic acid sequence encoding an engineered guide RNA;
- and
- a second promoter sequence operably linked to the second sequence,
- wherein the engineered guide RNA comprises:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUGAAAGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 7), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), and 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117);
- and
- 5′-AUGCAAC-3′.
52 more elements
A method of editing a target nucleic acid in a cell, comprising:
- delivering, into the cell, a Cas12f1 protein or a nucleic acid encoding the Cas12f1 protein, and an engineered guide RNA or a nucleic acid encoding the engineered guide RNA,
- which allows a CRISPR/Cas12f1 complex to be formed in the cell,
- wherein the CRISPR/Cas12f1 complex is capable of editing the target nucleic acid, and
- the engineered guide RNA comprises:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUGAAAGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 7), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), and 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117);
- and
- 5′-AUGCAAC-3′.
50 more elements
A method of editing a target nucleic acid in a cell, comprising:
- delivering, into the cell, a Cas12f1 protein or a nucleic acid encoding the same, and an engineered guide RNA or a nucleic acid encoding the same,
- which allows a CRISPR/Cas12f1 complex to be formed in the cell wherein the CRISPR/Cas12f1 complex is capable of editing the target nucleic acid, and
- the engineered guide RNA comprises:
- an engineered scaffold region, and
- a spacer,
- wherein the engineered scaffold region and the spacer are sequentially linked to each other in a 5′ to 3′ direction,
- the spacer comprises 10 nucleotides to 50 nucleotides and has a sequence complementary to a target sequence,
- a sequence of the engineered scaffold region is different from 5′-CUUCACUGAUAAAGUGGAGAACCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUA GAACUUGAGUGAAGGUGGGCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUC UUCGGAAAGUAACCCUCGAAACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAAGA AAGUUGCAGAACCCGAAUAGACGAAUGAAGGAAUGCAAC-3′ (SEQ ID NO:
- 315), and
- the sequence of the engineered scaffold region is such that the following sequences are sequentially linked to each other in a 5′ to 3′ direction:
- a sequence selected from the group consisting of 5′-A-3′, 5′-AA-3′, 5′-GAA-3′, 5′-AGAA-3′, 5′-GAGAA-3′, 5′-GGAGAA-3′, 5′-UGGAGAA-3′, 5′-GUGGAGAA-3′, 5′-AGUGGAGAA-3′, 5′-AAGUGGAGAA-3′ (SEQ ID NO:
- 16), 5′-AAAGUGGAGAA-3′ (SEQ ID NO:
- 17), 5′-UAAAGUGGAGAA-3′ (SEQ ID NO:
- 18), 5′-AUAAAGUGGAGAA-3′ (SEQ ID NO:
- 19), 5′-GAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 20), 5′-UGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 21), 5′-CUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 22), 5′-ACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 23), 5′-CACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 24), 5′-UCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 25), 5′-UUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 26), and 5′-CUUCACUGAUAAAGUGGAGAA-3′ (SEQ ID NO:
- 9);
- a sequence selected from the group consisting of 5′-CCGCUUCACUUAGAGUGAAGGUGG-3′ (SEQ ID NO:
- 430), 5′-CCGCUUCACCUUAGGAGUGAAGGUGG-3′ (SEQ ID NO:
- 431), 5′-CCGCUUCACCAUUAGUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 38), 5′-CCGCUUCACCAAUUAGUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 39), 5′-CCGCUUCACCAAAUUAGCUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 40), 5′-CCGCUUCACCAAAAUUAGACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 41), 5′-CCGCUUCACCAAAAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 42), 5′-CCGCUUCACCAAAAGCUUAGGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 43), 5′-CCGCUUCACCAAAAGCUUUAGAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 44), 5′-CCGCUUCACCAAAAGCUGUUAGUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 45), 5′-CCGCUUCACCAAAAGCUGUUAGUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 46), 5′-CCGCUUCACCAAAAGCUGUUUAGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 47), 5′-CCGCUUCACCAAAAGCUGUCUUAGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 48), 5′-CCGCUUCACCAAAAGCUGUCCUUAGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 49), and 5′-CCGCUUCACCAAAAGCUGUCCCUUAGGGGAUUAGAACUUGAGUGAAGGUGG-3′ (SEQ ID NO:
- 10);
- 5′-GCUGCUUGCAUCAGCCUAAUGUCGAGAAGUGCUUUCUUCGGAAAGUAACCCUCGA-3′ (SEQ ID NO:
- 11);
- a sequence selected from the group consisting of 5′-AACAAAGAAAGGA-3′ (SEQ ID NO:
- 110), 5′-AACAAAUGAAAAGGA-3′ (SEQ ID NO:
- 111), 5′-AACAAAUUGAAAAAGGA-3′ (SEQ ID NO:
- 112), 5′-AACAAAUUCGAAAGAAGGA-3′ (SEQ ID NO:
- 113), 5′-AACAAAUUCAGAAAUGAAGGA-3′ (SEQ ID NO:
- 114), 5′-AACAAAUUCAUGAAAAUGAAGGA-3′ (SEQ ID NO:
- 115), 5′-AACAAAUUCAUUGAAAAAUGAAGGA-3′ (SEQ ID NO:
- 116), 5′-AACAAAUUCAUUUGAAAGAAUGAAGGA-3′ (SEQ ID NO:
- 117), 5′-AACAAAUUCAUUUUGAAACGAAUGAAGGA-3′ (SEQ ID NO:
- 293), 5′-AACAAAUUCAUUUUUGAAAACGAAUGAAGGA-3′ (SEQ ID NO:
- 294), 5′-AACAAAUUCAUUUUUCGAAAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 295), 5′-AACAAAUUCAUUUUUCCGAAAAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 296), 5′-AACAAAUUCAUUUUUCCUGAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 297), 5′-AACAAAUUCAUUUUUCCUCGAAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 298), 5′-AACAAAUUCAUUUUUCCUCUGAAAAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 299), 5′-AACAAAUUCAUUUUUCCUCUCGAAAGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 300), 5′-AACAAAUUCAUUUUUCCUCUCCGAAACGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 301), 5′-AACAAAUUCAUUUUUCCUCUCCAGAAACCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 302), 5′-AACAAAUUCAUUUUUCCUCUCCAAGAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 303), 5′-AACAAAUUCAUUUUUCCUCUCCAAUGAAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 304), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUGAAAAACCCGAAUAGACGAAUGAAGGA-3′ (SEQ ID NO:
- 305), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCGAAAGAACCCGAAUAGACGAAUGAAGG A-3′ (SEQ ID NO:
- 306), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGAAAAGAACCCGAAUAGACGAAUGAA GGA-3′ (SEQ ID NO:
- 307), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGGAAACAGAACCCGAAUAGACGAAUG AAGGA-3′ (SEQ ID NO:
- 308), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCGAAAGCAGAACCCGAAUAGACGAAU GAAGGA-3′ (SEQ ID NO:
- 309), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCAGAAAUGCAGAACCCGAAUAGACGA AUGAAGGA-3′ (SEQ ID NO:
- 310), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACGAAAUUGCAGAACCCGAAUAGAC GAAUGAAGGA-3′ (SEQ ID NO:
- 311), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAGAAAGUUGCAGAACCCGAAUAG ACGAAUGAAGGA-3′ (SEQ ID NO:
- 312), 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAGAAAUUGCAGAACCCGAAUAGA CGAAUGAAGGA-3′ (SEQ ID NO:
- 313), and 5′-AACAAAUUCAUUUUUCCUCUCCAAUUCUGCACAAGAAAGUUGCAGAACCCGAAUA GACGAAUGAAGGA-3′ (SEQ ID NO:
- 314);
- and
- 5′-AUGCAAC-3′.
71 more elements
Dependencies are inferred from the claim text.