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T7 RNA polymerase engineering and RNA amplification

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
71677
Program Year/Program:
2004 / STTR
Agency Tracking Number:
GM072412
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
AMBION DIAGNOSTICS, INC.
Asuragen, Inc. 2150 WOODWARD ST AUSTIN, TX -
View profile »
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No
 
Phase 1
Fiscal Year: 2004
Title: T7 RNA polymerase engineering and RNA amplification
Agency: HHS
Contract: 1R41GM072412-01
Award Amount: $178,517.00
 

Abstract:

DESCRIPTION (provided by applicant): Microarrays are powerful tools that can impact new diagnostic procedures and expedite the drug development process. A significant but largely unrealized application is gene expression analysis from RNAlimited samples, such as blood, needle biopsies, laser capture microdissection (LCM) samples, and even single cells. The most widely used T7 RNA polymerase-based amplification method currenly lacks the necessary sophistication to meet these emerging needs. To overcome these limitations, we propose to increase the efficiency of T7 RNA amplification using an innovative protein engineering approach. Through combined rational design and random mutagenesis, mutant library selection and screening, we aim to identify a hyperactive T7 RNA polymerase that is 4-5 times more kinetically proficient than the current enzyme. We will expand this novel approach in phase II to include the discovery of mutant T7 polymerase enzymes that are both more thermostable (and thus extend high efficiency transcription for longer times, creating more product) and can incorporate biotinylated and Cy-modified nucleotides 5- to 10-fold more efficiently. Taken together, these improvements will lower the demand for input RNA by approximately 20-fold, and reduce the expense of modified nucleotides by as much as an order of magnitude.

Principal Investigator:

Liangjing Chen
5126510200
LCHEN@AMBION.COM

Business Contact:

Rigo Vallejo
5126510200
RVALLEJO@AMBION.COM
Small Business Information at Submission:

AMBION, INC.
2130 WOODWARD STREET AUSTIN, TX 78744

EIN/Tax ID: 742515119
DUNS: N/A
Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No
Research Institution Information:
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
UNIVERSITY OF TEXAS HLTH SCI CTR
Austin, TX 77225
RI Type: Nonprofit college or university