Multi-gene expression profile

• US 7,049,102 B1
• Filed: 11/15/2000
• Issued: 05/23/2006
• Est. Priority Date: 09/22/1989
• Status: Expired due to Fees

• 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: Eberwine, James H., Von Zastrow, Mark E., Barchas, Jack D., Van Gelder, Russell N.
• Primary Examiner(s): Ketter, James
• Assistant Examiner(s): KATCHEVES, KONSTANTINA T
• Application Number:
  US09/713,545
• Time in Patent Office: 2,015 Days
• Field of Search: 536/23.1, 536/24.1, 435/91.1
• US Class Current: 435/91.1
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• 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 2531/143 Promoter based amplification, e.g. NASBA, 3SR, TAS
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis

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

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

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

6. The multi-gene expression profile of claim 1, wherein said sample is a mammalian cell.

7. The multi-gene expression profile of claim 6, wherein said sample is a cell from brain, spleen, bone, heart, vascular tissue, lung, kidney, liver, pituitary, endocrine gland, lymph node, or tumor.

8. The multi-gene expression profile of claim 6, wherein said sample is a blood cell.

9. The multi-gene expression profile of claim 6, wherein same sample is a neural cell.

10. The multi-gene expression profile of claim 6, wherein said sample is a single cell.

4. The multi-gene expression profile of 1, wherein said amplified specific nucleic acid messages are hybridized to a hybridizing target.

5. The multi-gene profile of claim 4, wherein said amplified specific nucleic acid messages are hybridized to a hybridizing target by northern or Southern blot.

11. A multi-gene expression profile of a sample comprising a collection of linearly amplified specific nucleic acid messages, wherein said amplified specific nucleic acid messages have been amplified simultaneously with an RNA polymerase and with a single primer linked to an RNA polymerase promoter.

12. The multi-gene expression profile of claim 11, wherein said amplified specific nucleic acid messages are hybridized to a hybridizing target.