Methods for profiling mRNA expression in neurites

• US 5,723,290 A
• Filed: 11/03/1994
• Issued: 03/03/1998
• Est. Priority Date: 11/03/1994
• Status: Expired due to Term

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Eberwine, James, Miyashiro, Kevin, Dichter, Marc
• Primary Examiner(s): Myers, Carla J.
• Application Number:
  US08/334,254
• Time in Patent Office: 1,216 Days
• Field of Search: 435/6, 435/91.2, 435/91.21, 435/91.51, 536/23.5, 536/24.31, 935/17, 935/18, 935/78, 935/77
• US Class Current: 435/6.16
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

A method of identifying neurite cDNA clones by determining and comparing mRNA expression in selected neurites is provided. cDNA clones identified by this method are also provided. In addition, methods of profiling mRNA expression and diagnosing and treating conditions associated with a pattern of mRNA expression by determining an mRNA expression profile are provided.

Claims

1. A method of identifying neurite cDNAs comprising:

(a) converting an mRNA population in selected neurites into cDNA;

(b) determining mRNA expression in the selected neurites;

(c) comparing the relative levels of mRNA expression; and

(d) identifying neurite cDNAs based on the level of mRNA expression.

2. The method of claim 1 wherein the mRNA expression is determined by an aRNA amplification procedure.

3. A method of profiling mRNA expression in a selected neurite comprising:

(a) converting an mRNA population in a selected neurite into cDNA;

(b) making the cDNA double stranded;

(c) linearly amplifying the double stranded cDNA into aRNA; and

(d) using the aRNA as a probe in reverse phase Northern analysis to produce an mRNA expression profile of the selected neurite.

4. The method of claim 3 wherein the mRNA population in the selected neurite is converted into cDNA using an oligo-dT-T7 primer.

5. The method of claim 3 wherein the double stranded cDNA is linearly amplified into aRNA using T7 RNA polymerase.