US 11,702,686 B1
Compositions of matter for detection assays
Anurup Ganguli, San Diego, CA (US); Ariana Mostafa, San Diego, CA (US); Jacob Berger, San Diego, CA (US); Ashish Pandey, San Diego, CA (US); and Rashid Bashir, Urbana, IL (US)
Assigned to LabSimply, Inc., San Diego, CA (US); and The Board of Trustees of the University of Illinois, Urbana, IL (US)
Filed by LabSimply, Inc., San Diego, CA (US); and The Board of Trustees of the University of Illinois, Urbana, IL (US)
Filed on Feb. 6, 2023, as Appl. No. 18/106,420.
Application 18/106,420 is a continuation of application No. 17/861,208, filed on Jul. 9, 2022, granted, now 11,639,520.
Claims priority of provisional application 63/289,112, filed on Dec. 13, 2021.
Claims priority of provisional application 63/220,987, filed on Jul. 12, 2021.
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6823 (2018.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/682 (2018.01)
CPC C12Q 1/6823 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12Q 1/682 (2013.01); C12Q 1/6806 (2013.01); C12N 2310/20 (2017.05); C12N 2310/315 (2013.01); C12N 2310/321 (2013.01); C12N 2310/3231 (2013.01); C12N 2320/10 (2013.01)] 28 Claims
 
1. A composition of matter comprising a ribonucleoprotein (RNP) complex and a blocked nucleic acid molecule, wherein the blocked nucleic acid molecule is represented by Formula I, wherein Formula I in the 5′-to-3′ direction comprises:
A-(B-L)J-C-M-T-D;
wherein A is 0-15 nucleotides in length;
B is 4-12 nucleotides in length;
L is 3-25 nucleotides in length;
J is an integer between 1 and 10;
C is 4-15 nucleotides in length;
M is 1-25 nucleotides in length or is absent, wherein if M is absent then A-(B-L)j-C and T-D are separate nucleic acid strands;
T is 17-135 nucleotides in length and comprises at least 50% sequence complementarity to B and C;
D is 0-10 nucleotides in length and comprises at least 50% sequence complementarity to A; and
wherein the blocked nucleic acid molecule comprises a sequence complementary to a gRNA; and wherein the ribonucleoprotein complex comprises a nucleic acid-guided nuclease and the gRNA; and wherein the nucleic acid-guided nuclease exhibits cis-cleavage activity and trans-cleavage activity; wherein a Kd of the blocked nucleic acid molecules to the RNP is about 105-fold greater or more than the Kd of an unblocked nucleic acid molecule resulting from unblocking of the blocked nucleic acid molecules.