Akademik Veri Yönetim Sistemi
Tezin Türü: Yüksek Lisans
Tezin Yürütüldüğü Kurum: İstanbul Üniversitesi, Fen Fakültesi, Moleküler Biyoloji ve Genetik Bölümü, Türkiye
Tezin Dili: İngilizce
Öğrenci: İlknur YILMAZ
Asıl Danışman (Eş Danışmanlı Tezler İçin): Ahmed Sayed Ibrahım Aly
Eş Danışman: Bedia Palabıyık
Özet:
Ultraviolet
light damages DNA by converting any two adjacent thymines into a thymine dimer
that is potentially mutagenic, carcinogenic, or lethal to the organism. This
damage is repaired in all organisms (except mammals) by the enzyme Photolyase
[1,3]. Dr. Aziz SANCAR has won the Noble prize for the characterization of the
function of a homologue of the photolyase enzymes in higher eukaryotes. Here,
we are going to use the CRISPR-Cas9 technology to investigate function of DNA
Photolyase by targeted gene deletion, gene knockin-tagging and gene swap with
plants and yeast photolyases. We have designed the knock-out construct of our
study to contain the novel monomeric photo-convertible fluorescent protein
Eos4b. UV or blue light exposure of less than 1 minute transforms EOS4b protein
from bright green to bright red. Thus, we will be able to investigate the
effect of UV on knockout parasites using a UV sensitive marker. For the
Knockin-tagging study, we have designed the construct to tag the photolyase
protein at the C-terminus with the very bright and photostable NeonGreen or
Ruby3 fluorescent proteins, which contain the Strep Twin epitope tag. For the
gene swap design for to determine function of photolyase gene by gene swapping
of different photolyase and Cryptochrome genes found in other organisms into
the malaria parasite. In the future, knowing the detailed function of the
gene will lead to future studies to develop vaccine against malaria or to
develop an inhibitor for use in the treatment of patients