Increasing the sensitivity of a crispr-cas nucleic acid detection system
AssigneeNederlandse Organisatie voor toegepast-natuurwetenschappelijk onderzoek TNO
InventorHans Christiaan VAN LEEUWEN
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Abstract
The disclosure provides an amplification system for a nucleic acid detection system. This amplification system is particularly useful to increase the sensitivity of the nucleic acid detection system. Furthermore, the disclosure provides a method for detecting nucleic acids.
Claims22
A nucleic acid detection system, comprising a clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated nuclease (Cas) system (CRISPR-Cas system), and a signal amplification system, wherein:
- the CRISPR-Cas system comprises an effector protein and a target-specific guide RNA (gRNA) comprising a guide sequence capable of targeting the effector protein to a target sequence, and the effector protein exhibiting target-activated collateral nucleic acid cleavage activity;
- the signal amplification system comprises a partial CRISPR RNA (crRNA) recognition site-oligo (POISER-oligo) and a POISER-oligo-specific gRNA comprising a guide sequence capable of targeting an effector protein to the POISER-oligo further comprising a polynucleotide tail, and the effector protein exhibiting target-activated collateral nucleic acid cleavage activity;
- and
- a template-independent polymerase.
A signal amplification system for a nucleic acid detection system, comprising
- a partial crRNA recognition site-oligo (POISER-oligo),
- a POISER-oligo-specific guide RNA (gRNA) comprising a guide sequence capable of targeting an effector protein to the POISER-oligo further comprising a polynucleotide tail;
- and
- a template-independent polymerase.
Dependencies are inferred from the claim text.