Novel process, construct and conjugate for producing multiple nucleic acid copies

• US 20090068731A1
• Filed: 06/01/2007
• Published: 03/12/2009
• Est. Priority Date: 01/13/1994
• Status: Abandoned Application

• Current Assignee: Enzo Biochem Incorporated
• Original Assignee: Enzo Biochem Incorporated
• Inventors: Stavrianopoulos, Jannis G., Donegan, James J., Rabbani, Elazar, Engelhardt, Dean L.
• Application Number:
  US11/809,452
• Publication Number:
  US 20090068731A1
• Time in Patent Office: Days
• US Class Current: 435/320.100
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6853 using modified primers or templates
• CPC Class Codes: C12Q 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 1/6865 Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
• CPC Class Codes: C12Q 2521/119 RNA polymerase
• CPC Class Codes: C12Q 2525/203 incorporating a composite nucleic acid containing a polypeptide sequence other than PNA

Abstract

This invention provides inter alia an in vitro process for producing multiple specific nucleic acid copies in which the copies are produced under isostatic conditions, by degradation of primer sequences from extended primers, thereby allowing subsequent priming events on the same template.

Claims

1. A promoter-independent non-naturally occurring nucleic acid construct which when present in a cell produces a nucleic acid without the use of any gene product coded by said construct.

2. The construct of claim 1 comprising double-stranded and single-stranded nucleic acid regions.

3. The construct of claim 1 wherein said nucleic acid comprises deoxyribonucleic acid, ribonucleic acid, a DNA:

RNA copolymer, or a polymer capable of hybridizing or forming a base-specific pairing complex and initiating nucleic acid polymerization.

4. The construct of claim 1 comprising at least one modified nucleotide or nucleotide analog.

5. The construct of claim 1 comprising at least one single-stranded region.

6. The construct of claim 5 wherein said single-stranded region comprises a bubble.

7. The construct of claim 6 wherein said bubble comprises at least one complementary sequence to a nucleic acid present in the cell.

8. The construct of claim 7 wherein said bubble comprises at least one polyT sequence.