Compositions and methods of use for small-molecule regulation of CRISPR-Cas9 activity using RNA aptamers
AssigneesThe Regents of the University of Colorado, a body corporate, Inscripta, Inc.
InventorsRobert Batey, Roman S. Iwasaki Cordero, Andrew Garst
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Abstract
Provided herein are single guide RNAs (sgRNAs) that comprise aptamer sequences and related compositions and methods. Also provided herein are methods of selecting inducible sgRNAs that comprise aptamer sequences.
Claims19
A guide RNA (sgRNA) comprising a small-molecule-binding aptamer sequence adjacent to a region comprising a 2×4 internal loop and a region comprising an upper stem, wherein the small-molecule-binding aptamer is a theophylline-binding aptamer and the theophylline-binding aptamer comprises at least 85% homology to a polynucleotide represented by the sequence 5′-AUACCAGCUUCGAAAGAAGCCCUUGGCAG-3′ (SEQ ID NO:
- 95).
A guide RNA (sgRNA) comprising a small-molecule-binding aptamer sequence adjacent to a region comprising a 2×4 internal loop and a region comprising an upper stem, wherein the region comprising the 2×4 internal loop and the upper stem comprises the sequence 5′-GNNNGGAUACCAGCUUCGAAAGAAGCCNUUGGCAGCCNNNNNC-3′ (SEQ ID NO:
- 169).
A guide RNA (sgRNA) comprising a small-molecule-binding aptamer sequence adjacent to a region comprising a 2×4 internal loop and a region comprising an upper stem, wherein the region comprising the 2×4 internal loop and the upper stem comprises a polynucleotide represented by SEQ ID NO:
- 100, UGAAGGAUACCAGCUUCGAAAGAAGCCNUUGGCAGCCGCAACA.
Dependencies are inferred from the claim text.