	US 12,319,963 B2
	Type V CRISPR/Cas effector proteins for cleaving ssDNAs and detecting target DNAs
Jennifer A. Doudna, Berkeley, CA (US); Janice S. Chen, Berkeley, CA (US); Lucas Benjamin Harrington, Berkeley, CA (US); and Enbo Ma, Moraga, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Filed by The Regents of the University of California, Oakland, CA (US)
Filed on Jul. 29, 2021, as Appl. No. 17/388,321.
Application 17/388,321 is a continuation of application No. 16/896,731, filed on Jun. 9, 2020, granted, now 11,118,224.
Application 16/896,731 is a continuation of application No. 16/262,257, filed on Jan. 30, 2019, abandoned.
Application 16/262,257 is a continuation of application No. 15/897,089, filed on Feb. 14, 2018, granted, now 10,253,365, issued on Apr. 9, 2019.
Claims priority of provisional application 62/626,593, filed on Feb. 5, 2018.
Claims priority of provisional application 62/590,106, filed on Nov. 22, 2017.
Prior Publication US 2021/0388437 A1, Dec. 16, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/6876 (2018.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12Q 1/6823 (2018.01)

CPC C12Q 1/6876 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12Q 1/6823 (2013.01); C12N 2310/20 (2017.05)]

21 Claims

1. A method of assaying for a target nucleic acid in a sample, the method comprising:

a. contacting the sample to a volume comprising:

i. a type V CRISPR/Cas effector protein,

ii. a guide nucleic acid comprising a nucleotide sequence that hybridizes to a nucleic acid segment of the target nucleic acid or an amplification or transcription product thereof, wherein the target nucleic acid or the amplification or transcription product thereof is DNA, and

iii. a detector DNA molecule; and

b. assaying for a signal produced by cleavage of the detector DNA molecule by the type V CRISPR/Cas effector protein upon hybridization of the guide nucleic acid to the nucleic acid segment of the target nucleic acid or the amplification or transcription product thereof, wherein assaying for the signal comprises measuring a change in the signal over time, wherein measuring the change in signal over time comprises measuring the change in signal in real-time, the method further comprising determining an amount of the target nucleic acid.