20.500.12556/RUL-172880 Tehnologija CRISPR-Cas9 in njena uporaba CRISPR-Cas9 technology and its applications CRISPR-Cas sistem, ki je naravno prisoten pri določenih bakterijah in arhejah, predstavlja pomembno orodje v biotehnologiji. Boljše razumevanje in nadaljnji razvoj te tehnologije bosta omogočila napredek na številnih področjih, kot so gensko inženirstvo, zdravljenje bolezni, kmetijstvo, prehranska industrija, ekologija, farmacija in mnoga druga. Najpogosteje uporabljan sistem temelji na endonukleazi Cas9 in njenih derivatih (dCas9, nCas9). Poznamo več aplikacij CRISPR-Cas9. Med najpomembnejše v genskem inženirstvu sodijo klasični sistem CRISPR-Cas9 in modificirani sistemi, ki omogočajo regulacijo izražanja genov (CRISPR-dCas9), urejanje posameznih nukleotidov in epigenetske modifikacije. Med sodobnejše pristope spada urejanje »prime«, ki poleg zamenjave posameznih nukleotidov omogoča tudi uvedbo manjših insercij ali delecij na tarčna mesta. Slednji predstavlja velik preboj na tem področju, saj so z baznimi urejevalci in sistemom urejanja »prime« uspeli preseči omejitve klasičnega CRISPR-Cas9 sistema. Kljub velikemu potencialu se tehnologija sooča s kar nekaj pomanjkljivostmi, med katerimi so najpomembnejša potencialni izventarčni učinki. Ker pri uporabi in vivo izventarčni učinki niso sprejemljivi, je ključnega pomena, da jih poskušamo odpraviti. Poleg tehničnih omejitev tehnologije pa se pojavljajo tudi etična vprašanja, zlasti v povezavi s preurejanjem genoma na zarodkih in spolnih celicah, kar bi lahko pomenilo prenos sprememb na naslednje generacije in možne nepredvidljive posledice. The CRISPR-Cas system, which occurs naturally in certain bacteria and archaea, is an extremely important genetic tool in biotechnology. A deeper understanding and further development of this technology will enable advances in numerous fields such as genetic engineering, disease treatment, agriculture, food industry, ecology, pharmacy and many others. The most commonly used system is based on the Cas9 endonuclease and its derivatives (dCas9, nCas9). There are several applications of the CRISPR-Cas9 system. Among the most important for genetic engineering are the classic CRISPR-Cas9 system and the modified systems that enable gene regulation, base editing and epigenetic modifications. A more recent approach is prime editing, which enables the insertion or deletion of small sequences at target sites in addition to editing of individual nucleotides. The latter represents a major breakthrough in this field, as base editors and prime editing systems have successfully overcome the limitations of classic CRISPR-Cas9. Despite its enormous potential, the technology faces several limitations, the most critical of which are off-target effects. As these are unacceptable in in vivo applications, it is imperative that they are eliminated. In addition to these limitations, there are also ethical concerns, particularly in relation to genome editing of embryos and germ cells, which could lead to the transmission of changes to future generations and thus to unpredictable outcomes. biotehnologija genski inženiring CRISPR-Cas9 dCas9 urejanje posameznih baz urejanje »prime« biotechnology genetic engineering CRISPR-Cas9 dCas9 base editing prime editing true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2025-09-12 07:16:32 2025-09-12 07:16:37 2025-10-27 03:28:41 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 263635 248765187 51004.pdf 51004.pdf 1 7150C582BA5B88F016D28DD01CDB9982 07f0e89659241cedb80724fbbcd151e34ce09a683634336ca34650bcda32c1ee 8016422d-8f97-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=217014 Biotehniška fakulteta 0 0 0