The focus of this research will be to evaluate vertically-aligned carbon nanofiber (VACNF) arrays as a parallel interface to deliver macromolecules, specifically different DNAs, in a spatially resolved manner to cellular matrices. Our nanofiber-based approach exploits a universal delivery mechanism, microinjection, that has been demonstrated as effective for all cell types (mammalian, plant, bacterial). However, unlike conventional microinjection, which is a labor intensive and slow serial method for single cell delivery, vast arrays of VACNFs can provide for massively parallel gene delivery to cellular matrices. Spatially resolved delivery will allow the production of arrays of genetically manipulated live cells that can be used as a powerful discovery tool. Examples of applications and future R01 efforts that can be supported by successful demonstration of this tool include the real-time functional characterization of large ensembles of expressed gene products, the evaluation of specificity of candidate drugs on libraries of over-expressing transformed cells, and loss-of-function analyses in cell groups receiving or producing interfering RNAs. Towards this goal, this effort will evaluate the potential of nanofiber-mediated gene delivery, with the specific target of spatially resolved material delivery to cellular matrices for live-cell arraying. In this R21 effort we will: 1. Investigate strategies for temporarily and permanently immobilizing different DNAs in discrete patterns to localized regions of periodic arrays of vertically-aligned carbon nanofibers grown on planar substrates 2. Incorporate these DNA-modified arrays into cellular matrices in a parallel 'microinjection'-based scheme and subsequently quantify the effectiveness of plasmid delivery and expression within targeted cells. 3. Initially evaluate nanofiber-mediated methods for live cell microarraying of diverse cell types; including both mammalian and yeast cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21EB004066-01
Application #
6809535
Study Section
Special Emphasis Panel (ZRG1-GDD (01))
Program Officer
Moy, Peter
Project Start
2004-08-01
Project End
2006-07-31
Budget Start
2004-08-01
Budget End
2005-07-31
Support Year
1
Fiscal Year
2004
Total Cost
$169,213
Indirect Cost
Name
University of Tennessee Knoxville
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
003387891
City
Knoxville
State
TN
Country
United States
Zip Code
37996
Mann, David G J; McKnight, Timothy E; Melechko, Anatoli V et al. (2007) Quantitative analysis of EDC-condensed DNA on vertically aligned carbon nanofiber gene delivery arrays. Biotechnol Bioeng 97:680-8