Development of a tool for tracking H3K27 tri-methylation

H3K27 trimethylation is a post-translational modification known to play a role in faithful chromosome segregation via its interaction with TopoIIα. The ability to observe the spatiotemporal dynamics of this modification and how it interacts with TopoIIα has thus far been a challenge. Here, we have utilized a method involving an inducible fluorescent modification-specific intracellular antibody (mintbody) that reversibly binds to post-translational modifications to tag the H3K27 trimethylation. Four cell lines were created using a mintbody including super folded GFP that can be induced with treatment with doxycycline. Different numbers of guide RNAs were also used in an attempt to increase CRISPR/Cas9 efficiency. These cell lines were screened and analyzed for future use in live cell imaging and in vivo experiments concerning the interplay of H3K27 trimethylation, TopoIIα, and chromosome segregation.

Grantor dc:publisher

University of Kansas

Year dc:date.issued

2024

Author dc:creator

• Bertolino, Morgan

Advisor dc:contributor.advisor

• Azuma, Yoshiaki

• Cellular biology
• Molecular biology
• CRISPR
• H3K27me3
• histone posttranslational modifications
• mintbody

Statement dc:rights

• This item is protected by copyright and unless otherwise specified the copyright of this thesis/dissertation is held by the author.

Language dc:language.iso

en

Dc Identifier Other

http://dissertations.umi.com/ku:19743

OAI identifier oai:identifier

oai:kuscholarworks.ku.edu:1808/36672