Možnosti reprogramiranja in transdiferenciacije celic z uporabo aktivatorjev CRISPR.

Doneva, Angela (author), Ogorevc, Jernej (mentor)

Reprogramiranje celic in transdiferenciacija pomenita spreminjanje identitete in funkcije celic. V diplomski nalogi sva pregledala literaturo in preučila možnosti za reprogramiranje in transdiferenciacijo celic z uporabo aktivatorjev vezanih na encim Cas, ki je del CRISPR/Cas ribonukleoproteinskega kompleksa. Sistem CRISPR/Cas je zelo popularno orodje za urejanje genoma, uporaben pa je tudi za uravnavanje izražanja različnih genov. Temelji na vzpostavitvi kompleksa vodilne RNA (sgRNA – single guide RNA), ki določa tarčno regijo, in proteina Cas. Ob določenih modifikacijah je sistem uporaben tudi za drugačno aplikativno rabo. Ena od takšnih modifikacij je zamenjava proteina Cas9 z mutirano obliko, ki nima endonukleazne aktivnosti (»dead« Cas9-dCas9). Dokazano je bilo, da fuzija transaktivacijskih domen na dCas9, ob vezavi na tarčna mesta v genomu, inducira izražanje tarčnih genov, s čimer lahko dosežemo reprogramiranje celic v pluripotentno stanje oziroma jih diferenciramo v želeni celični tip. V literaturi smo poiskali primere, kjer je bila tehnologija že uspešno uporabljena v ta namen in poskušali bomo ovrednotiti prednosti in slabosti regulacije genov, ki temelji na CRISPR/dCas9

• Language: Slovenian
• Keywords: CRISPR sistem, dCas9, CRISPRa, matične celice, iPSC-inducirane matične celice, reprogramiranje celic, transdiferenciacija
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [A. Doneva]
• Year: 2019
• PID: 20.500.12556/RUL-110439
• UDC: 602.641:602.9:611.018(043.2)
• COBISS.SI-ID: 9369977
• Publication date in RUL: 14.09.2019

DONEVA, Angela, 2019, Možnosti reprogramiranja in transdiferenciacije celic z uporabo aktivatorjev CRISPR. [online]. Bachelor’s thesis. A. Doneva. [Accessed 3 June 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=110439

Secondary language

• Language: English
• Title: Cell reprogramming and transdiferentiation with the use of CRISPR actvators

Abstract

Cell reprogramming and transdifferentiation mean changing the identity and function of cells. In the thesis we reviewed the literature and examined the possibilities for reprogramming and transdifferentiation of cells using activators bound to the Cas enzyme that is part of the CRISPR / Cas ribonucleoprotein complex. The CRISPR / Cas system is a very popular tool for gene editing. It is based on the establishment of ribonucleoprotein complex, consisting of RNA (sgRNA) which determines the target region, and the Cas protein. With certain modifications, the system can also be used for different applicative uses. One such modification is replacement of a Cas9 protein with a mutated form that lacks endonuclease activity ("dead" Cas9-dCas9). It has been shown that fusion of transactivation domains to dCas9, upon binding to target sites in the genome, induces the expression of target genes, which may be used for cell reprogramming or cell the aim of achieving reprogramming of cells in a pluripotent state or differentiation into the desired cell type. Literature mining revealed that the technology has been successfully used fort his purpose and could be useful in future regenerative medicine and biotechnology applications.

• Keywords: CRISPR system, dCas9, CRISPRa, stem cells, iPSC-induced stem cells, cell reprogramming, transdifferentiation