The purpose of this project is to test the possibility of developing TiO2 -biopolymer nanocomposites as new vehicles for biotechnology. We propose to use TiO2-DNA nanocomposites as inducible gene specific endonucleases, with allele-differentiating sequence specificity. This function of nanocomposites relies on (a) TiO2 nanoparticle dependent charge separation inducing DNA cleavage; and (b) sequence specificity of oligonucleotides attached to TiO2 nanoparticle.
The specific aims of this proposal are: 1. In vitro studies (outside cells) of the interaction between HSVtk specific nanocomposites and DNA and RNA specific for the HSVtk (Herpes Simplex Virus thymidine kinase) gene 2. Studies of the interaction between HSVtk specific nanocomposites and DNA specific for the HSVtk gene in cells in culture 3. Evaluation of mutations caused by interactions between HSVtk specific nanocomposites and HSVtk gene in cells in culture. We will study DNA scission following activation of single and multiple nanocomposites (with one or two different oligonucleotides attached per nanoparticle) annealed to the target sequence. The target sequence is Herpes simplex virus thymidine kinase gene (HSVtk). We chose this sequence to be our target because HSVtk acts as a suicide gene in cells treated with guanine analogues, and therefore mutagenesis of the HSVtk sequence provides cells with a selective advantage when guanine analogues are added into the cell medium. We believe that upon completion of these studies we will be able to link results of in vitro DNA scission assays with the spectra and frequency of mutations in cells in culture, use this knowledge to predict the behavior of nanocomposites inside cells, and associate the type of mutation that can be anticipated with different nanocomposite sequences and modes of nanocomposite activation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
1R01EB002100-01A2
Application #
6918175
Study Section
Special Emphasis Panel (ZRG1-BECM (01))
Program Officer
Moy, Peter
Project Start
2005-04-01
Project End
2009-01-31
Budget Start
2005-04-01
Budget End
2006-01-31
Support Year
1
Fiscal Year
2005
Total Cost
$300,713
Indirect Cost
Name
Northwestern University at Chicago
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
Brown, Koshonna; Thurn, Ted; Xin, Lun et al. (2018) Intracellular in situ labeling of TiO2 nanoparticles for fluorescence microscopy detection. Nano Res 11:464-476
Kurepa, Jasmina; Nakabayashi, Ryo; Paunesku, Tatjana et al. (2014) Direct isolation of flavonoids from plants using ultra-small anatase TiO? nanoparticles. Plant J 77:443-53
Bazak, Remon; Ressl, Jan; Raha, Sumita et al. (2013) Cytotoxicity and DNA cleavage with core-shell nanocomposites functionalized by a KH domain DNA binding peptide. Nanoscale 5:11394-9
Falaschetti, Christine A; Paunesku, Tatjana; Kurepa, Jasmina et al. (2013) Negatively charged metal oxide nanoparticles interact with the 20S proteasome and differentially modulate its biologic functional effects. ACS Nano 7:7759-72
Yuan, Ye; Chen, Si; Paunesku, Tatjana et al. (2013) Epidermal growth factor receptor targeted nuclear delivery and high-resolution whole cell X-ray imaging of Fe3O4@TiO2 nanoparticles in cancer cells. ACS Nano 7:10502-17
Yuan, Y; Chen, S; Gleber, S C et al. (2013) Mapping the subcellular localization of Fe3O4@TiO2 nanoparticles by X-ray Fluorescence Microscopy. J Phys Conf Ser 463:
Jensen, Mark P; Aryal, Baikuntha P; Gorman-Lewis, Drew et al. (2012) Submicron hard X-ray fluorescence imaging of synthetic elements. Anal Chim Acta 722:21-8
Paunesku, T; Wanzer, M B; Kirillova, E N et al. (2012) X-ray fluorescence microscopy for investigation of archival tissues. Health Phys 103:181-6
Arora, Hans C; Jensen, Mark P; Yuan, Ye et al. (2012) Nanocarriers enhance Doxorubicin uptake in drug-resistant ovarian cancer cells. Cancer Res 72:769-78
Yuan, Ye; Paunesku, Tatjana; Arora, Hans et al. (2011) Interrogation of EGFR Targeted Uptake of TiO2 Nanoconjugates by X-ray Fluorescence Microscopy. AIP Conf Proc 1365:423-426

Showing the most recent 10 out of 38 publications