วันพุธที่ 25 กุมภาพันธ์ พ.ศ. 2558

MINOR PROJECT

Pyrrolidinyl peptide nucleic acid internally-labeled with cyanine-styryl dye as a fluorescence DNA probe
My research question is to find excellent the environment-sensitive fluorescence labels, alkyne-modified cyanine-styryl derivatives (STR), which were labeled onto pyrrolidinyl peptide nucleic acid with D-proline/(1S,2S)-2-aminocyclopentanecarboxylic acid backbone (acpcPNA) via sequential reductive alkylation-Click strategy with the aim to create self-reporting PNA probes that exhibit fluorescence change in response to the correct DNA target. The styryl dye family has received increasing attention as a photostable DNA label that revealed the excellent colorimetric response in difference microenvironments.
Researchers who have looked at this subject are Wen-hai Zhan and Rubner. Both of studies focused on the synthesis and evaluation of styryl dyes capable of exhibiting turn-on fluorescence through sequence-specific interaction with nucleic acid in fluorescence spectroscopy and diagnostic application.
Zhan et al. (2005) argue that the 2-propynyl substituted hemicyanine dyes were synthesized in a facile route and showed good optical properties that have a strong absorbance as well as a large strokes shift of the fluorescence emission maxima. The terminal alkynyl group of dyes was successfully employed for bioconjugation through a Click strategy with azide-modified cowpea mosaic virus (CPMV).
Rubner et al. (2012) argue that the alkyne-modified indole-quinoline styryl dye (CyIQ) was synthesized and incorporated it into DNA via click chemistry. The CyIQ dye exhibit good brightness and give excellent photostability.
Debate centers on this basic issue of the fluorescent properties of cyanine-styryl dyes which exhibit good brightness. Thus, we are interested in synthesizing new clickable styryl dyes for attachment onto acpcPNAs with the aim to use them as DNA sensing probes.
My work will be closer to Rubner’s in the focus on the synthesis of the alkyne-modified styryl family for fluorescent labeling and incorporated into nucleic acid as fluorescent probes.
Hopefully my contribution will be to synthesize and incorporate into acpcPNA which discriminate between the complementary DNA and single mismatch DNA and apply in SNP detection for diagnosis.
Reference list
Zhan, W-h., Barnhill, H. N., Sivakumar, K., Tian, H., Wang, Q. (2005). Synthesis of hemicyanine dyes for ‘click’ bioconjugation. Tetrahedron Letters, 46, 1691-1695.

Rubner, M. M., Holzhauser, C., Bohlander, P. R., Wagenknecht, H-A. (2012). A “Clickable” Styryl Dye for Fluorescent DNA labeling by Excitron and Energy Transfer Interactions. Chemistry - A European Journal, 18, 1299-1302. 

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