PMID- 27583462
OWN - NLM
STAT- MEDLINE
DCOM- 20170712
LR  - 20181202
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 114
DP  - 2016 Aug 4
TI  - Observation and Quantification of Telomere and Repetitive Sequences Using 
      Fluorescence In Situ Hybridization (FISH) with PNA Probes in Caenorhabditis 
      elegans.
LID - 10.3791/54224 [doi]
LID - 54224
AB  - Telomere is a ribonucleoprotein structure that protects chromosomal ends from 
      aberrant fusion and degradation. Telomere length is maintained by telomerase or 
      an alternative pathway, known as alternative lengthening of telomeres (ALT)(1). 
      Recently, C. elegans has emerged as a multicellular model organism for the study 
      of telomere and ALT(2). Visualization of repetitive sequences in the genome is 
      critical in understanding the biology of telomeres. While telomere length can be 
      measured by telomere restriction fragment assay or quantitative PCR, these 
      methods only provide the averaged telomere length. On the contrary, fluorescence 
      in situ hybridization (FISH) can provide the information of the individual 
      telomeres in cells. Here, we provide protocols and representative results of the 
      method to determine telomere length of C. elegans by fluorescent in situ 
      hybridization. This method provides a simple, but powerful, in situ procedure 
      that does not cause noticeable damage to morphology. By using fluorescently 
      labeled peptide nucleic acid (PNA) and digoxigenin-dUTP-labeled probe, we were 
      able to visualize two different repetitive sequences: telomere repeats and 
      template of ALT (TALT) in C. elegans embryos and gonads.
FAU - Seo, Beomseok
AU  - Seo B
AD  - Institute of Molecular Biology and Genetics (IMBG), Seoul National University.
FAU - Lee, Junho
AU  - Lee J
AD  - Institute of Molecular Biology and Genetics (IMBG), Seoul National University; 
      Department of Biological Sciences, Seoul National University; Department of 
      Biophysics and Chemical Biology, Seoul National University; elegans@snu.ac.kr.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20160804
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (DNA Probes)
RN  - 0 (Peptide Nucleic Acids)
RN  - EC 2.7.7.49 (Telomerase)
SB  - IM
MH  - Animals
MH  - Caenorhabditis elegans/*genetics/metabolism
MH  - DNA Probes/chemistry/genetics
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Peptide Nucleic Acids/chemistry/genetics
MH  - Repetitive Sequences, Nucleic Acid
MH  - Telomerase/genetics/metabolism
MH  - Telomere/genetics/*metabolism
PMC - PMC5091719
EDAT- 2016/09/02 06:00
MHDA- 2017/07/14 06:00
PMCR- 2018/08/04
CRDT- 2016/09/02 06:00
PHST- 2016/09/02 06:00 [entrez]
PHST- 2016/09/02 06:00 [pubmed]
PHST- 2017/07/14 06:00 [medline]
PHST- 2018/08/04 00:00 [pmc-release]
AID - 54224 [pii]
AID - 10.3791/54224 [doi]
PST - epublish
SO  - J Vis Exp. 2016 Aug 4;(114):54224. doi: 10.3791/54224.