Visualization of long human telomere mimics by single-molecule fluorescence imaging.

J Phys Chem B 2008 Oct 26;112(42):13184-7. Epub 2008 Sep 26.

Study of long single-stranded telomeric DNA is important for a variety of basic science and biotechnological applications, yet few methods exist for synthesis and visualization of single copies of this DNA in solution at biologically relevant length scales necessary for assessment of heterogeneity in its structure and behavior. We have synthesized kilobase-long single-stranded human telomere mimics in situ by rolling circle replication (RCR) on a microscope coverslip surface and visualized individual strands by staining with SYBR Gold. Under buffer flow, differential extensibility and varying morphology of these long telomere-mimicking DNA sequences were observed at the single-molecule level in real time. Using this procedure, we detected striking differences in the extensibility of individual RCR products based on the human G-rich telomeric sequence in the presence and absence of short, complementary single-stranded oligonucleotides. We also apply this new mode of single-stranded DNA characterization to probe the interaction of kilobase-length telomere mimics with the small-molecule G-quadruplex-binding agent TMPyP4.

Download full-text PDF

Source
http://dx.doi.org/10.1021/jp806696uDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2688642PMC
October 2008
5 Reads

Publication Analysis

Top Keywords

telomere mimics
12
human telomere
8
varying morphology
4
morphology long
4
extensibility varying
4
buffer flow
4
flow differential
4
differential extensibility
4
long telomere-mimicking
4
dna sequences
4
level real
4
real time
4
single-molecule level
4
observed single-molecule
4
gold buffer
4
sequences observed
4
telomere-mimicking dna
4
staining sybr
4
rolling circle
4
circle replication
4

Similar Publications