Multi-genes expression profile

• US 6,291,170 B1
• Filed: 04/05/1999
• Issued: 09/18/2001
• Est. Priority Date: 09/22/1989
• Status: Expired due to Term

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford University)
• Inventors: Von Zastrow, Mark E., Barchas, Jack D., Van Gelder, Russell N., Eberwine, James D.
• Primary Examiner(s): Schwartzman, Robert A.
• Assistant Examiner(s): SANDALS, WILLIAM O
• Application Number:
  US09/286,533
• Time in Patent Office: 897 Days
• Field of Search: 536/23.1, 536/24.31, 435/6, 435/320.1
• US Class Current: 435/6.14
• CPC Class Codes: C12N 15/1096 cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-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]

Abstract

This invention relates to the use of promoters for ribonucleic acid amplification and other genetic manipulations. Processes are provided wherein complementary deoxyribonucleic acid (cDNA) is synthesized from a ribonucleic acid (RNA) sequence using a complementary primer linked to an RNA polymerase promoter region complement and then anti-sense RNA (aRNA) is transcribed from the cDNA by introducing an RNA polymerase capable of binding to the promoter region. Additional processes using the resulting aRNA are also described.

Claims

1. A multi-gene expression profile of a sample comprising

a hybridizing target;

and

a hybridizing probe comprising a collection of amplified specific nucleic acid messages, which amplified specific nucleic acid messages have been amplified in vitro simultaneously with RNA polymerase and a single primer linked to a RNA polymerase promoter, wherein said amplified specific nucleic acid messages each have a relative abundance which qualitatively reflects the relative representation of specific nucleic acid messages within the sample;

whereby the collection of amplified specific nucleic acid messages is a probe hybridizing to the target to form a multi-gene expression profile.

2. The multi-gene expression profile of claim 1, wherein the amplified specific nucleic acid messages comprise aRNA.

3. The multi-gene expression profile of claim 1, wherein the amplified specific nucleic acid messages comprise cDNA.

5. The multi-gene expression profile of claim 1, wherein the single cell is a neural cell.

6. The multi-gene expression profile of claim 5 wherein the relative level of representation varies as a function of arousal state, behavior, drug treatment or development.

4. A multi-gene expression profile of a single cell comprising

a collection of amplified specific nucleic acid messages, which amplified specific nucleic acid messages have amplified in vitro simultaneously with an RNA polymerase and a single primer linked to an RNA polymerase promoter, wherein each of said amplified specific nucleic acid messages has a relative abundance which qualitatively reflects the relative abundance of the specific nucleic acid message within the sample;

and

a hybridizing target;

whereby the collection of amplified specific nucleic acid messages hybridizes to the target to form the multi-gene expression profile.