	US 11,884,921 B2
	Signal boost cascade assay
Anurup Ganguli, San Diego, CA (US); Ashish Pandey, San Diego, CA (US); Ariana Mostafa, San Diego, CA (US); and Jacob Berger, San Diego, CA (US)
Assigned to VedaBio, Inc., San Diego, CA (US)
Filed by LabSimply, Inc., San Diego, CA (US)
Filed on Dec. 9, 2022, as Appl. No. 18/078,821.
Claims priority of provisional application 63/397,785, filed on Aug. 12, 2022.
Claims priority of provisional application 63/395,394, filed on Aug. 5, 2022.
Claims priority of provisional application 63/359,183, filed on Jul. 7, 2022.
Claims priority of provisional application 63/289,112, filed on Dec. 13, 2021.
Prior Publication US 2023/0193273 A1, Jun. 22, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/113 (2010.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01)

CPC C12N 15/113 (2013.01) [C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 2310/20 (2017.05)]

5 Claims

1. A method for preventing unwinding of blocked nucleic acid molecules in the presence of an RNP comprising the steps of:

providing blocked nucleic acid molecules;

providing ribonucleoprotein complexes comprising a Cas12a nucleic acid-guided nuclease that exhibits both cis- and trans-cleavage activity upon activation and a gRNA that recognizes an unblocked nucleic acid molecule resulting from trans-cleavage of the blocked nucleic acid molecules; and

engineering the nucleic acid-guided nuclease used in the ribonucleoprotein complex to result in a variant nucleic acid-guided nuclease where single stranded DNA is cleaved faster than double stranded DNA is cleaved.