Primjena CRISPR/Cas9 tehnologije u biljaka i implikacije na zakonodavnu regulativu

undergraduate thesis

• Title (english): Application of CRISPR/Cas9 technology in plants and its implication on legislative regulation
• Author: Mirela Kubat
• 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

Posljednjih nekoliko godina svjedočimo revoluciji u biologiji do koje je došlo otkrićem nove kategorije nukleaza koje omogućuju precizno uređivanje genoma. Kao nukleaze u takvom sustavu za uređivanje genoma, do sada su se koristile zinc finger nukleaze (ZFN), efektorske nukleaze nalik transkripcijskim aktivatorima (TALEN), RNA-navođene nukleaze (RGN) iz CRISPR i CRISPR/Cas sustava. Zbog jednostavnosti kojom se može prilagoditi za uređivanje genoma, CRISPR/Cas9 sustav je ubrzo postao najrašireniji alat. Sam CRISPR/Cas sustav je dio adaptivnog imuniteta bakterija i arheja, koji ih štiti od stranih nukleinskih kiselina tako što ih specifično cijepa. Dva su osnovna dijela, sgRNA koja služi kao vodeća i Cas9 nukleaza. Takav sustav uvodi dvolančani lom (DSB) u slijed DNA. Popravak loma staničnim sustavom popravka kao rezultat ima ciljane modifikacije genoma. CRISPR/Cas tehnologija značajno povećava mogućnost poboljšanja odlika usjeva u usporedbi s konvencionalnim pristupima razmnožavanju. Sve spomenute nukleaze uvode male insercije i delecije (indele) ili supstitucije na ciljnim mjestima na kromosomima koje se ne može razlikovati od prirodno nastalih genetičkih varijacija. Biljke uređene CRISPR/Cas tehnologijom bi se mogle komercijalizirati mimo postojeće regulative, ali će se vjerojatno još uvijek smatrati GMO od strane regulatornih autoriteta u određenim državama, što bi moglo smanjiti korištenje programiblinih nukleaza u biotehnologiji biljaka i agrikulturi. Mogućnosti otvara DNA-neovisno uređivanje genoma, s prethodno sastavljenim ribonukleoproteinima. U biljkama ne ostaje rekombinantna DNA, te bi se bijke proizvedene DNA-neovisno mogle isključiti iz važećih GMO regulativa, omogućujući masovno korištenje RNA-vođenog uređivanja genoma u biljnoj biotehnologiji i agrikulturi.

Abstract (english)

In the last couple of years, we are witnessing a revolution in biology, brought by discovery of nucleases which enable high precision genome editing. Till this day the nucleases used were zinc finger nucleases (ZFN), transcription activator like effector nucleases, and RNA-guided nucleases from CRISPR/Cas systems. Due to it's simplicity, CRISPR/Cas system shortly became the most common used tool. Originaly, the CRISPR/Cas system is found in bacteria and archea, as a part of adaptive immune system, which protects them from exogenous nucleic acids, by sequence-specific cliving. In the system, an sgRNAis present, which guides the nuclease, Cas9. This system makes a double strand DNA break. Site- specific modifications result from fixing the DSB by endogenous systems. CRISPR/Cas technology increases the possibility of enhancing crop traits by a great deal in comparison with classic crop breeding tehniques. Programmable nucleases introduce small indels or substitutions which can not be distinguished from naturally occuring genetic variations. Plants edited by CRISPR/Cas could be commercialized bypassing the existing regulative, but will probably still be considered GMO in certain states, which could reduce usage of programmable nucleases in plant biotechnology and agriculture. DNA-free genome editing opens some possibilities. In plants that are edited with preassembled RNPs, there isn't any recombinant DNA left. These plants could be excluded from existing GMO regulatives, and open doors into mass usage of RNA-guided genome editing in plant biotechnology and agriculture.

• Keywords: CRISPR/Cas9 tehnologija; biljke
• Language: croatian
• URN:NBN: urn:nbn:hr:217:318520
• 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 12:38:06