Transgenic mice for study of the CDK8/19 cyclin-dependent kinase kinase-independent mechanisms of action

Stavskaya NI1, Ilchuk LA2, Okulova YuD2, Kubekina MV2, Varlamova EA2, Silaeva YY1, Bruter AV2
About authors

1 Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia

2 Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia

Correspondence should be addressed: Leonid A. Ilchuk
Prospekt Mira, 124, Moscow, 129164, Russia; moc.liamg@21kuhcel

About paper

Funding: RSF grant, project № 22-15-00227.

Author contribution: Stavskaya NI ― experimental procedure, working with animals; Ilchuk LA ― manuscript writing, design of genotyping systems, data analysis; Okulova YuD ― working with embryos; Kubekina MV ― preparation of genetically engineered construct, experimental procedure; Varlamova EA ― experimental procedure; Silaeva YuYu ― literature analysis, study planning; Bruter AV ― literature analysis, study planning, data analysis and interpretation, manuscript editing.

Compliance with ethical standards: the study was approved by the Ethics Commitee of the Institute of Gene Biology RAS (protocol No. 1 of 10 November 2021) and conducted in full compliance with the Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes.

Received: 2022-11-22 Accepted: 2022-12-19 Published online: 2022-12-28

The CDK8 cyclin-dependent transcription-associated kinase and its less studied paralog, CDK19, regulate the expression of the dependant genes via several mechanisms. CDK8/19 can directly phosphorylate some  transcription factors (ICN, STAT1), but at the same time these kinases being a component of the mediator complex regulate transcrition via interaction with chromatin in the promoter and enhancer regions of appropriate genes. Recently the papers have appeared showing that CDK8/19 has kinase-independent mechanisms of action through comparison of the effects of the kinase activity genetic inactivation and chemical inhibition. The study was aimed to generate transgenic mice capable of the induced and tissue-specific expression of the kinase-negative (showing no phosphorylation activity) form of CDK8, CDK8 (D173A), which could be later used to study the CDK8 kinase-independent mechanisms of action in vivo. We obtained four F0 transgenic animals by microinjection of linear DNA into the pronucleus, two of these animals became the ancestors of two distinct lines. The copy number of the integrated construct was measured for all F0 and the lines generated. This model may be used to study the kinase-independent properties of the CDK8/19 proteins.

Keywords: transgenesis, transcription regulation, Cdk8, mediator kinase, kinase-independent functions