Kvantitativne analize telomera

undergraduate thesis

• Title (english): Quantitative telomere analysis
• Author: Doris Zorić
• Mentor: Ivica Rubelj (mentor)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2015, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Duljina telomera, kao i dinamika njihovog skraćivanja, pokazatelj je dobi i replikativnog potencijala stanice te je pouzdan biomarker za procjenu dugovječnosti i praćenje bolesti povezanih sa starenjem. Sve češća upotreba rezultata mjerenja duljine telomera u dijagnostici zahtijeva točnu i preciznu kvantitativnu analizu i statističku obradu podataka. Kvantitativna analiza telomera danas se provodi s različitim spektrom metoda, od kojih je uvjerljivo najkorištenija i najbolje interpretirana analiza terminalnih restrikcijskih fragmenata (TRF) metodom hibridizacije po Southernu. Glavni nedostatak ove analize jest činjenica da njome ne saznajemo stvarnu duljinu telomera, već prosječnu duljinu terminalnih fragmenata (telomera s pridruženom subtelomernom regijom) na razini stanice ili populacije. Ostale često korištene metode poput kvantitativnog PCR-a ili Q-PNA-FISH-a imaju nekoliko prednosti u odnosu na hibridizaciju po Southernu, no zbog velike varijacije u rezultatima ili nemogućnosti usporedbe rezultata i dalje su manje pouzdane. Zbog proširenog znanja o naglom skraćivanju pojedinih telomera kao uzroku senescencije, teži se individualnijem pristupu i usavršavanju metoda za analizu duljine pojedinih telomera te metoda za brzu kliničku primjenu (primjerice, HPA). Otegotna okolnost kvantitativne analize telomera jest njihovo svojstvo heterogenosti po veličini uvjetovano zbivanjima na molekularnoj razini DNA, stanice i stanične populacije, koja uz metodološka ogrančenja producira i heterogenost rezultata. Kvantitativna analiza, stoga, zatijeva nužno statističku obradu i ispravnu interpretaciju podataka. Točni rezultati analize telomera tada mogu biti primjenjivi u dijagnostici preuranjenog staničnog starenja, ranom otkrivanju bolesti i aproksimaciji maksimalne životne dobi te dati informacije o biološkim fenomenima za daljnja fundamentalna znanstvena istraživanja.

Abstract (english)

Telomere length, just like the dynamics of its shortening, is indicator of the cell age and its replicative potential but also a reliable biomarker for lifespan approximation and the monitoring of aging-related diseases. The results provided by telomere length measurement depend upon the correct and precise quantitative and statistical analysis. Quantitative analysis of telomeres is determine by several methods, among which the most commonly used and most reliable one is terminal restriction fragment (TRF) length analysis by Southern blot. The main disadvantage of the aforementioned method is the fact that it doesn't show the actual telomere length, but the mean terminal fragment with subtelomeric tails for given cell population. Other often applied methods such as the quantitative PCR or Q-PNA-FISH have some advantages compared to Southern blot. Yet, due to significant variation of the results or imposibility of their comparison among different studies these methods are not first choice in fundamental research. Wider knowledge about the single telomere shortening to critical level, when senescence is induced, resulted in increasing effort in creating a more individual approach in telomere measurement. Still, the aggravating circumstance of quantitative telomere analysis is the teleomere property of intrinsic heterogeneity. It is caused by molecular processes at the DNA level, cell level and cell population level. In addition, the methodological limitations add to increased heterogenity of results. Therefore, quantitative analysis unquestionably needs to be accompanied by a statistical analysis and an accurate interpretation of results. Precise telomere analysis results can be applied in cell senescense, studies of various diseases and population lifespan distribution.

• Keywords: telomere; kvantitativne analize telomera
• Keywords (english): telomeres; quantitative telomere analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:449152
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) molekularne biologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-12-19 09:41:52