diplomsko delo
Matic Kostanjšek (Author), Irena Golinar (Mentor), Irena Golinar (Thesis defence commission member), Polona Jamnik (Thesis defence commission member), Mojca Narat (Thesis defence commission member)

Abstract

Sistem CRISPR/Cas9 je bakterijski način obrambe proti virusom. V zadnjih letih je postal najbolj uporabljena metoda za spreminjanje genomov. Sistem z enojno vodečo (sgRNA) specifično prepoznava točno določena zaporedja na genomu, kamor se nato veže protein Cas9 z nukleazno aktivnostjo. Zaradi enostavnosti spreminjanja zaporedja sgRNA in s tem tarče v genomu, ima sistem veliko aplikacij na številnih področij, od spreminjanja genoma rastlin za produkcijo terapevtskih proteinov, mikrobne biotehnologije, sintetične biologije, do direktnih kliničnih aplikacij pri ljudeh. Poleg dobro poznanih aplikacij sistema, s katerimi se genom spremeni, pa se je razvilo tudi ogromno drugih aplikacij. Sistem z deaktiviranim proteinom Cas9 se tako danes uporablja tudi za regulacijo prepisovanja genov ali spreminjanje epigenoma. Kljub širokemu spektru obstoječih aplikacij pa obstajajo omejitve, kot so netarčne mutacije ali težave z dostavo, ki zavirajo še širšo uporabo sistema. Pojavilo se je že veliko rešitev za te ovire. V zadnjem času je bilo izdelanih nekaj izboljšanih proteinov Cas9, prav tako pa je bilo razvitih več novih sistemov, s pomočjo katerih bi lahko težave omejili. Poleg tega je zelo pomembno predvideti in kasneje tudi validirati potencialne neželene učinke v obliki netarčnih mutacij.

Keywords

biotehnologija;CRISPR/Cas9;genski inženiring;dCas9;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [M. Kostanjšek]
UDC: 601.4:577.21:602.62/.64:602.68(043.2)
COBISS: 9234297 Link will open in a new window
Views: 827
Downloads: 257
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Other data

Secondary language: English
Secondary title: Applications of CRISPR/Cas9 system, current limitations and future outlook
Secondary abstract: CRISPR/Cas9 is a bacterial immune system against invading viruses. Because altering the sgRNA sequence, which binds to the target in the genome is so facile, CRISPR/Cas9 has become a method of choice in the fields such as genetic engineering, microbial biotechnology or synthetic biology. Furthermore, the system has a great potential in clinical applications for correcting disease-causing genes in somatic cells. The potential of CRISPR/Cas9 extends further than gene editing via NHEJ or HDR. Applications of deactivated Cas9 protein with a fused effector domain have seen a surge in popularity in recent years. With them, it is possible to alter gene regulation, cause epigenetic remodeling or visualize certain loci of interest. Although there is a wide range of potential uses of the system, it is currently coping with some limitations. Such limitations are off-target mutations or problems with delivery. Recently, new solutions have emerged for those challenges, such as engineered Cas9 proteins, modifications in sgRNA and some new systems for higher specificity. It is crucial to determine potential off target sites in both in silico and in vivo or in vitro systems.
Secondary keywords: biotechnology;genetic engineering;
Type (COBISS): Bachelor thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: VIII, 22 str.
ID: 11149683