Sintetski genomi

undergraduate thesis

• Title (english): Synthetic genomes
• Author: Eva Pavlinek
• Mentor: Nenad Malenica (mentor)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2017, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Napredak biotehnologije doveo je do jednostavne sinteze kratkih oligonukleotida željenog slijeda u DNA sintetizatorima. Već godinama se razvijaju metode kojima bi se takvi fragmenti spajali u veće, sve do cjelovitih sintetskih genoma. Standardne metode sastavljanja pomoću restrikcijskih enzima i ligaze imaju već odužu povijest pa tako i ograničenja. Iako je temeljena na tom principu, metoda BioBricks je danas široko primijenjena zahvaljujući modifikacijama. Danas se rutinski koriste modernije metode koje ne zahtijevaju restrikcijske enzime, već se temelje na preklapajućim homolognim sljedovima. Specifične egzonukelaze, polimeraze i ligaze su sposobne prerađivati takve sljedove te od njih sastavljati veće fragmente DNA. Primjeri takvih metoda su metode sastavljanja DNA In-Fusion te Gibson Assembly.
Navedene metode se koriste u proučavanju strukture i funkcije genoma, kao i mogućnosti unaprjeđenja samih metoda. Najistaknutiji uspješni projekt sinteze genoma je sintetski genom
Synthia, gdje je uspješno stvorena bakterijska stanica potpuno kontrolirana kemijski sintetiziranim genomom. Daljnja istraživanja su pružila mnoge spoznaje o esencijalnim genima te mogućnosti modifikacije sintetskih genoma.
Aktualni projekt The Genome Project – Write istražuje mogućnosti sinteze većih eukariotskih genoma. Do sada su najznačajniji uspjesi postignuti s kvaščevim sintetskim genomom, a daljnji planovi stavljaju naglasak na sintezu ljudskog genoma.

Abstract (english)

The progress of biotechnology has led to a simple synthesis of short oligonucleotides of the desired sequence in DNA synthesizers. Many methods have been developed to assemble them into larger fragments, even to complete synthetic genomes. Although standard assembly methods based on restriction enzymes and ligases have been used for a long time, they are limited for today’s purposes. Such method, BioBrick assembly is widely applied thanks to modifications. Currently developed are modern methods which do not require restriction enzymes, but are based on overlapping homologous ends. Specific exonucleases, polymerases and ligases are capable of processing such sequences and assembling them into larger DNA fragments. Examples of such methods are In-Fusion and Gibson Assembly.
Mentioned methods are used in studying the structure and function of the genome, as well as the possibilities of improving the methods themselves. So far, the most prominent genome synthesis project is synthetic genome Synthia, where researchers created a bacterial cell completely controlled by chemically synthesized genome. Further research has provided many insights into the nature of genomes and the potential for their modification.
Current project The Genome Project - Write explores possibilities for synthesis of larger eukaryotic genomes. Up to now, the most significant successes have been achieved with the yeast synthetic genome, and further plans emphasize the synthesis of the human genome.

• Keywords: sintetski genomi
• Language: croatian
• URN:NBN: urn:nbn:hr:217:679655
• 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: 2017-10-23 12:53:39