# UNIVERSAL SYSTEM FOR DIRECTED NUCLEIC ACID CLEAVAGE

> **NIH NIH R43** · THIRD WAVE TECHNOLOGIES · 1994 · $75,000

## Abstract

We propose to develop a novel system for directed nucleic acid cleavage.
This system is based upon our discovery that the 5' nucleases of several
thermostable DNA polymerases can be directed to cleave as site-specific
endonucleases. This system will be thermostable, specific, efficient and
easily automatable, and will find broad application in the fields of
nucleic acids-based research and diagnostics. In Phase I we will seek to
overcome the most substantial limitation of this technology, its
inefficiency at cleaving long 5' arms. We will examine the effects of
reaction constituents and conditions on the efficiency of cleavage of a
well-characterized substrate, performing a comprehensive examination of
the effects of different salts, divalent cations, denaturants, buffer pH
values, helicases and temperatures. We will also use mutagenesis to alter
the protein and influence its interaction with the substrate structure.
During Phase II we will optimize the required substrate complex formations
and nuclease interactions to develop this technology into a generic,
directed nucleic acid cleavage system for both single and double stranded
nucleic acids.

## Key facts

- **NIH application ID:** 2190398
- **Project number:** 1R43GM051704-01
- **Recipient organization:** THIRD WAVE TECHNOLOGIES
- **Principal Investigator:** MARY A BROW
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $75,000
- **Award type:** 1
- **Project period:** 1994-07-01 → 1994-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2190398

## Citation

> US National Institutes of Health, RePORTER application 2190398, UNIVERSAL SYSTEM FOR DIRECTED NUCLEIC ACID CLEAVAGE (1R43GM051704-01). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/2190398. Licensed CC0.

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