Function of Zinc finger protein 644 (Zfp644) in mouse organism.

• Název práce v češtině: Funkce Zinc-finger proteinu 644 (Zfp644) v myším organismu.
• Název v anglickém jazyce: Function of Zinc finger protein 644 (Zfp644) in mouse organism.
• Klíčová slova: zinc-finger, transgenní myš, mutace, epigenetika, Zfp644, ZNF644, WIZ, G9a, GLP, metylace, zrak, plodnost, vyvoj,
• Klíčová slova anglicky: Zfp644, ZNF644, WIZ, G9a, GLP, histone methylation, transcription factor, vision, fertility, development, transgenic mouse, mutation, epigenetics
• Akademický rok vypsání: 2016/2017
• Typ práce: disertační práce
• Jazyk práce: angličtina
• Ústav: Katedra genetiky a mikrobiologie (31-140)
• Vedoucí / školitel: doc. RNDr. Radislav Sedláček, Ph.D.
• Řešitel: skrytý - zadáno vedoucím/školitelem
• Datum přihlášení: 21.10.2016
• Datum zadání: 21.10.2016
• Datum odevzdání elektronické podoby: 27.10.2021
• Datum proběhlé obhajoby: 26.01.2022
• Oponenti: doc. RNDr. Kateřina Komrsková, Ph.D.
• Konzultanti: Mgr. Jan Procházka, Ph.D.

Předběžná náplň práce v anglickém jazyce

The Zfp644 gene is encoding for a zinc finger protein in GWAS study associated as potential genetic factor of inherited myopia. In 2014, the Radislav Sedlacek team developed a novel mouse model using TALEN technology targeting the Zfp644 gene (ZNF644 in human) in order to reveal molecular mechanism of Zfp644 action. Initial phenotyping research was done in department of transgenic models of diseases in 2015 with Zfp644 KO mice. By systematic description of phenotype, it was identified slower growth, defects in spermatids maturation and slightly abdominal obesity. Also Zfp644 KO mice showed behavioral changes. In vivo micro CT scans (skeleton morphology, body composition) and histopathology screen showed no significant alternation either in body alternation or composition. Interestingly, preliminary results show strong expression of Zfp644 in testis, suggesting that phenotype in spermatid maturation can be primary effect of mutation.
In 2015 Di Giacomo et al. (2014) published that Zfp644 is a binding protein that has a repressive transcription mechanism in the germ line. WIZ (a zinc finger protein similar to Zfp644) and GLP complex is directed to specific genomic loci which contains DNA binding sites for WIZ and Zfp644 to silence gene expression. Affected target genes are CWH43, DIP2C and ROCK1. This two zinc finger proteins can also deposit the G9a/GLP complex on chromatin independently. Examples are for Zfp644 binding in the promotors for CACNA2D1 and ANKRD26P1, or for WIZ which binds to the promoters of PARD3 and ABCA13. Also Bian (et al. 2015) published that Zfp644 regulate the G9a/GLP complex for gene repression. WIZ contents 12 zinc finger motifs and interact with the catalytic domain of G9a, while Zfp644 contents 8 zinc finger motifs and N – terminus of Zfp644 interacts with TAD of GLP.
The main aim of the PhD thesis is to find the molecular function of Zinc finger protein 644 (Zfp644).
To achieve this, my research project will be focus on :

1. Systematic expression mapping of Zfp644 expression during embryogenesis and adult tissues by whole-mount in situ hybridization and RT-PCR.
2. Unbias phenotyping screens with use of CCP infrastructure and transgenic unit (fertility phenotyping).
3. The robust trascriptomics analysis (RNASeq) to compare testis transcriptomes in Zfp644 null and WT mice.
4. ChiP seq in order to identify binding sites and sequences recognized by Zfp644.
5. Analyze genome methylation status in testis of Zfp644 KO mice.
6. With biochemistry approaches (co-immuno precipitation) followed by mass spec analysis I will make attempt to identify binding partners in presumptive methylation complex.