公开(公告)号：US20250146089A1

公开(公告)日：2025-05-08

申请号：US18936289

申请日：2024-11-04

Applicant: The Trustees of Princeton University

Inventor： Caitlin Lamb ,  Arend Jan Wouter te Velthuis ,  Cameron Myhrvold

IPC: C12Q1/70 ,  C12N9/22 ,  C12N15/11 ,  C12Q1/6818

Abstract: Disclosed is a method for quantitative detection of a small, aberrant viral nucleic acid. The method includes combining a detection mixture with a sample, and allowing a reaction with the test mixture to begin. The sample includes an RNA sample. The detection mixture includes a Cas13 enzyme, a reporter, a sequence-specific CRISPR RNA (crRNA), and a buffer. The method includes determining a quantitative value representative of the reporter in the test mixture, where the quantitative value is a measure of an amount of target RNA present in the RNA sample.

公开(公告)号：US20250115951A1

公开(公告)日：2025-04-10

申请号：US18908108

申请日：2024-10-07

Applicant: The Trustees of Princeton University

Inventor： Cameron Myhrvold ,  Benjamin Larsen ,  Ofer Kimchi ,  Owen Dunkley ,  Aartjan te Velthuis

IPC: C12Q1/6823 ,  C12N9/22 ,  C12N15/11 ,  C12Q1/683 ,  C12Q1/6832

Abstract: Disclosed are techniques for enhanced nucleic acid detection using blocked or partially blocked crRNAs or target RNAs, and a composition of matter (detection reactions consisting of occluded crRNAs or target RNAs). Here, the mechanism of Cas 13 protein activation in response to RNA structure perturbations was systematically probed using a massively multiplexed screen. It was found that there are two distinct sequence-independent modes by which secondary structure affects Cas13 activity: structure in the protospacer region competes with the crRNA and can be disrupted via a strand-displacement mechanism, while structure in the region 3′ to the protospacer has an allosteric inhibitory effect. The kinetic nature of the strand displacement process was leveraged to improve Cas13-based RNA detection and enhance mismatch discrimination by up to 50-fold, enabling sequence-agnostic mutation identification at low (

公开(公告)号：US20250066842A1

公开(公告)日：2025-02-27

申请号：US18814007

申请日：2024-08-23

Applicant: MASSACHUSETTS INSTITUTE OF TECHNOLOGY ,  PRESIDENT AND FELLOWS OF HARVARD COLLEGE ,  THE TRUSTEES OF PRINCETON UNIVERSITY

Inventor： Pardis Sabeti ,  Cameron Myhrvold ,  Sameed Siddiqui

IPC: C12Q1/682 ,  C12N9/02 ,  C12N9/22 ,  C12N15/11

Abstract: Cas-cleavable reporter systems, CRISPR-Cas systems thereof, and methods of use thereof in CRISPR-Cas based diagnostics. Cas-cleavable reporter systems may generate a luminescent, fluorescent, or other detectable signal upon Cas-collateral cleavage of one or more reporter system components. CRISPR-Cas systems may comprise a Cas protein having collateral cleavage activity, guide molecules, and the Cas-cleavable reporter system. Cas-cleavable reporter systems may be a split luciferase reporter system, at least one element of which is bound to beads. For multiplexing, CRISPR-Cas systems may comprise a Cas-cleavable quenched reporter system, optionally a Cas-cleavable quenched fluorescent reporter system, a Cas protein having collateral cleavage activity, and target-specific guide molecules attached to color-coded beads. CRISPR-Cas systems may further comprise amplification reagents. Methods may apply said CRISPR-Cas systems to flow cell, well-plate, or other devices, and may measure detectable signals by microscopic, plate reader, or other methods.