US 12,416,008 B2
Compositions and methods of use for small-molecule regulation of CRISPR-Cas9 activity using RNA aptamers
Robert Batey, Boulder, CO (US); Roman S. Iwasaki Cordero, Boulder, CO (US); and Andrew Garst, Boulder, CO (US)
Assigned to The Regents of the University of Colorado, a body corporate, Denver, CO (US); and Inscripta Inc., Boulder, CO (US)
Filed by THE REGENTS OF THE UNIVERSITY OF COLORADO, A BODY CORPORATE, Denver, CO (US); and INSCRIPTA INC., Boulder, CO (US)
Filed on Jul. 14, 2021, as Appl. No. 17/375,808.
Application 17/375,808 is a continuation of application No. PCT/US2020/013718, filed on Jan. 15, 2020.
Claims priority of provisional application 62/793,748, filed on Jan. 17, 2019.
Prior Publication US 2022/0042019 A1, Feb. 10, 2022
Int. Cl. C12N 15/115 (2010.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01)
CPC C12N 15/115 (2013.01) [C12N 9/22 (2013.01); C12N 15/111 (2013.01); C12N 2310/16 (2013.01); C12N 2310/20 (2017.05)] 19 Claims
 
1. 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).