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T7 RNA polymerase engineering and RNA amplification

Award Information
Agency: Department of Health and Human Services
Branch: National Institutes of Health
Contract: 2R42GM072412-02
Agency Tracking Number: GM072412
Amount: $672,066.00
Phase: Phase II
Program: STTR
Solicitation Topic Code: N/A
Solicitation Number: N/A
Timeline
Solicitation Year: 2007
Award Year: 2007
Award Start Date (Proposal Award Date): N/A
Award End Date (Contract End Date): N/A
Small Business Information
ACCACIA INTERNATIONAL, INC. 2113 WELLS BRANCH PKWY, STE 6900
AUSTIN, TX 78728
United States
DUNS: 084994735
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 ANDREW ELLINGTON
 (512) 232-3426
 andy.ellington@mail.utexas.edu
Business Contact
 EULA DEQUEIRA
Phone: (512) 784-1861
Email: BIOAID@AUSTIN.RR.COM
Research Institution
 UNIVERSITY OF TEXAS AT AUSTIN
 
UNIVERSITY OF TEXAS AUSTIN PO BOX 7726
AUSTIN, TX 78713 1980
United States

 Nonprofit College or University
Abstract

DESCRIPTION (provided by applicant): Expanding use of microarray systems for global gene expression profiling is leading biological research, providing diagnostic and prognostic value to physicians and facilitating target discovery during drug and vaccine development. There is strong interest in extending this microarray capability to more RNA-limited material to the point that a single cell can be profiled with confidence. The most widely used method for preparing samples for microarray analysis, which uses cDNA synthesis from mRNA followed by T7 RNA polymerase-based amplification, currently lacks the necessary sensitivity to meet these emerging needs. To overcome these methodological limitations, we propose to increase the final amplified RNA yield by 1) improving the efficiency of T7 RNA polymerase using structure-based engineering with directed evolution of function, 2) by optimizing the design of the promoter-template construct for specific amplification needs and 3) by re-standardization of cDNA synthesis for lower RNA inputs. The combined improvements are expected to lower the cell-limited RNA sample requirement by 100-fold compared to current, commercially available kits. This will enable microarray gene analysis with as little as 1 ng of total RNA with a single round of amplification or microarray profiling from a single cell (-10 pg of total RNA) after two rounds of amplification.

* Information listed above is at the time of submission. *

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