Use of CRISPR-Cas9 in genetic modification of poplar for improvment of phytoremediation abilities

Naše dejavnosti na Zemlji so v antropocenskem obdobju, v katerem smo okolje zastrupili s težkimi kovinami, herbicidi, eksplozivi in z drugimi kemikalijami, močno škodile rastlinam in živalim. Znanstveniki so začeli razvijati različne vrste remediacije šele takrat, ko smo se posledic naših dejanj zavedli, vendar je bila večina teh remediacij zelo draga in zelo zahtevna za izvedbo. Najcenejša, vendar časovno najbolj zahtevna metoda je fitoremediacija, torej remediacija okolja s pomočjo rastlin. Izmed vseh vrst rastlin je za tovrstni postopek zaradi svoje odpornosti in sposobnosti za hitro adaptacijo na različna okolja najbolj primeren topol. Njegova edina težava je njegova počasna presnova, ki pa je lahko za potrebe fitoremediacije ustrezno spremenjena s pomočjo genetske manipulacije. Na področju genetske manipulacije je trenutno zelo priljubljena metoda CRISPR. Cilj te diplomske naloge je predstaviti uporabo metode za utišanje spolnih genov pri topolih, s čimer bi pridobili sterilne topole, ki bodo lahko uporabni kljub omejitvam, ki so določene z zakonom za uporabo genetsko spremenjenih rastlin. Poleg tega je namen naloge podati ideje za izboljšanje metode CRISPR pri vstavljanju genov, ker je prav ta postopek izjemno pomemben pri ustvarjanju topolov z izboljšanimi fitoremediacijskimi sposobnostmi. Pri topolih so bili z drugimi metodami za genetsko modifikacijo že testirani različni geni, ki izboljšujejo fitoremediacijo. Vendar obstaja tudi veliko genov, katerih ogromni potencial je dokazan pri manjših rastlinah, katere bi jih bilo v prihodnosti dobro testirati tudi na topolih. Vstavljanje različnih fitoremediacijskih genov z metodo CRISPR bi zato lahko bilo zelo učinkovit način za remediacijo onesnaženih območij.Our activities on this planet in the anthropocene have had a negative influence on plants and animals due to the contamination of the environment with heavy metals, herbicides, explosives and other chemicals. Scientists started developing different methods for remediation only once we became aware of the consequences of our actions, though most of them are expensive and technically demanding. The most affordable, but time-consuming method is remediation by plants, i.e. phytoremediation. Among all the different species of plants the most suitable for phytoremediation is poplar, due to its resistance and ability to adapt to different environments. The problem is that poplar trees have slow metabolism, but for phytoremediation purposes this can be solved with genetic engineering. CRISPR technology is currently very popular in the field of genetic engineering. Our goal is to use this technology to suppress the floral genes in poplars, which would result in sterile poplars that could be used also in cases where existing laws prohibit the use of genetically modified plants. We also want to come up with ideas for improving the CRISPR method for gene insertion since this process is extremely important in creating poplars with improved phytoremediation ability. Various genes that improve phytoremediation have already been tested in poplars by using other methods for genetic modification. However, there are also many such genes that have their great potential proven in smaller plants, which should be tested on poplars in the future as well. Insertion of various genes for phytoremediation by the CRISPR method could be a very effective method for the remediation of contaminated zones