Knockdown von Runx2 durch RNA-Interference in chondrogenen Progenitor-Zellen

Knockdown of Runx2 with RNA-interference in chondrogenic progenitor cells

by Moritz Klein
Doctoral thesis
Date of Examination:2014-03-26
Date of issue:2014-03-26
Advisor:Prof. Dr. Nicolai Miosge
Referee:Prof. Dr. Nicolai Miosge
Referee:Prof. Dr. Dr. Karl Günter Wiese
Persistent Address: http://dx.doi.org/10.53846/goediss-4428

 

 

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Abstract

English

The regeneration of diseased hyaline cartilage continues to be a great challenge because its self renewal capacity is limited.  In repair tissue from human articular cartilage during the late stages of osteoarthritis chondrogenic progenitor cells (CPC`s) were found. These cells showed stemm cell capacity such as multipotency and migratory activity. The Transkriptionfaktor Runt-related transkription Factor 2 (Runx2) is an importend regulator of Osteogenesis and is also expressed in CPC`s.  We tried to generate a Knockdown of Runx2 in CPC`S with the help of an lentiviral Vektor and RNA-Interference. We also investigated the influence of Runx2 knockdown to the chondrogenic transkriptionfactor SRY-related HMG-box gene 9 (Sox9). Our results showed that the transfection of CPC`s with the i-lenti-GFP expression Vector (Biocat) with Amaxa Nukleofection leads to transient transfektion. The Knockdown of Runx2 could be showed to 96 h after transfektion with Polymerase chain  reaction (RT-PCR) and to 48 h in the western blot. We also could show a changing in sox9 expression in the transfected CPC`s.
Keywords: Runx2; Sox9; Osteoarthritis; RNA-interference; chondrogenic progenitorcells; Nucleofaction
 
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