公开(公告)号：US11608519B2

公开(公告)日：2023-03-21

申请号：US16049460

申请日：2018-07-30

申请人： TOKITAE LLC

发明人： Ted A. Baughman ,  Damian Madan ,  Eric Nalefski ,  Anne-Laure M. Le Ny

IPC分类号： C12Q1/6832 ,  C12N9/22 ,  C12N15/11

摘要： Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target DNA sequences from various sources using a Cas12a protein. When activated by binding a target DNA sequence, the Cas12a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target DNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target DNA sources, for the presence or absence of target DNA sequences. Compositions and kits, useful in practicing these methods, for example detecting a target DNA in a biological sample, are also described.

公开(公告)号：US20220106647A1

公开(公告)日：2022-04-07

申请号：US17430235

申请日：2020-02-07

申请人： Tokitae LLC

发明人： Ted Baughman ,  Anne-Laure M. Le Ny ,  Philip Leung ,  Damian Madan ,  Eric Nalefski

IPC分类号： C12Q1/6897 ,  C12N9/22 ,  C12N15/11

摘要： Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target nucleic acid sequences from various sources in a solution-based system. When activated by binding a target nucleic acid sequence, the Cas protein cleaves a tether separating a reporter molecule from a capture moiety. The capture moiety can then be used to remove, localize, or sequester uncleaved molecule containing intact tethers. In some embodiments, the systems, methods, and devices may include a filter, a membrane, or other molecules that may help to separate the tethered and untethered reporter molecules and/or capture the reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target nucleic acid, in some cases, without needing to amplify the target sequences, and without the need for sophisticated or expensive laboratory equipment. These devices and methods may be used to assay a wide variety of samples and target nucleic acid sources, for the presence or absence of a specific target sequences. Compositions and kits, useful in practicing these methods, for example detecting a target RNA or DNA in a biological sample, are also described.

公开(公告)号：US20200032325A1

公开(公告)日：2020-01-30

申请号：US16049460

申请日：2018-07-30

申请人： TOKITAE LLC

发明人： Ted A. Baughman ,  Damian Madan ,  Eric Nalefski ,  Anne-Laure M. Le Ny

IPC分类号： C12Q1/6832 ,  C12N9/22 ,  C12N15/11

摘要： Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded and double-stranded target DNA sequences from various sources using a programmed Cas12a protein. When activated by binding a target DNA sequence, the Cas12a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target DNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target DNA sources, for the presence or absence of a specific target DNA sequences. Compositions and kits, useful in practicing these methods, for example detecting a target DNA in a biological sample, are also described.

公开(公告)号：US20200032324A1

公开(公告)日：2020-01-30

申请号：US16049457

申请日：2018-07-30

申请人： TOKITAE LLC ,  THE REGENTS OF THE UNIVERSITY OF CALIFORNIA

发明人： Ted A. Baughman ,  James Cate ,  Jennifer Doudna ,  Gavin John Knott ,  Damian Madan ,  Eric Nalefski ,  Anne-Laure M. Le Ny ,  Brittney Thornton

IPC分类号： C12Q1/6832 ,  C12N9/22 ,  C12N15/11

摘要： Embodiments disclosed herein include devices, methods, and systems for direct, selective, and sensitive detection of single-stranded target RNA sequences from various sources using a programmed Cas13a protein. When activated by binding a target RNA sequence, the Cas13a cleaves a tether releasing a reporter molecule that may then be detected. In some embodiments, the systems, methods, and devices may include a filter or membrane that may help to separate the tethered and untethered reporter molecules. These devices, systems, and techniques allow a user to rapidly process samples that may contain the target RNA, without needing to amplify the target sequences. These devices and methods may be used to assay a wide variety of samples and target RNA sources, for the presence or absence of a specific target RNA sequence. Compositions and kits, useful in practicing these methods, for example detecting a target RNA in a biological sample, are also described.