T7 RNA polymerase engineering and RNA amplification

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$1,348,691.00
Award Year:
2007
Program:
STTR
Phase:
Phase II
Contract:
2R42GM072412-02
Award Id:
71677
Agency Tracking Number:
GM072412
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
ACCACIA INTERNATIONAL, INC., 2113 WELLS BRANCH PKWY, STE 6900, AUSTIN, TX, 78728
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
084994735
Principal Investigator:
ANDREWELLINGTON
(512) 232-3426
ANDY.ELLINGTON@MAIL.UTEXAS.EDU
Business Contact:
EULADEQUEIRA
() -
Research Institute:
UNIVERSITY OF TEXAS AT AUSTIN

UNIVERSITY OF TEXAS AUSTIN
PO BOX 7726
AUSTIN, TX, 78713 1980

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 use s 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) i mproving 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 synthes is 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 wit h 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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