Development of a molecular tool to study the function of anti-tumor genes.

ARJONA, ANDREA; AGÜERO, TRISTAN

Tucuman

Congreso; XXXIX ANNUAL SCIENTIFIC MEETING TUCUMAN BIOLOGY ASSOCIATION; 2023

Complex genetic changes occur during embryonic development that in a strictly coordinated manner promote the correct formation of an embryo. Studying these genetic mechanisms is a experimental challenge. Notably, tumor suppressor proteins are expressed during embryonic development. These proteins regulate different cell growth mechanisms and alterations in their function promote tumors development and malformations. The Bap1 protein is involved in tumor suppression mechanisms and is mutated in some of the most lethal cancers, such as melanomas. Bap1 selectively associates with the transcription factor Yin Yang1 (YY1) and both are part of an epigenetic repressor complex involved in transcriptional regulation. To carry out this work, we designed a molecular tool capable of selectively knock down the function of Yy1 in the context of a developing embryo. This tool, based on genome editing techniques, allows the change of gene function in vivo. To perform this loss-of-function strategy we analyzed the Xenopus Yy1 gene sequence and then compared it with its orthologous gene in humans using bioinformatic methods. We designed specific guide RNAs against Yy1 and together with a Cas13 nuclease we were able to degrade the messenger RNA of the target gene. Yy1 loss-of-function produces changes in neural plate morphology, a reduction of neural crest and cranial sensory placodes cells that will give rise the eye, and also defects in the craniofacial morphology of developing embryos. The use of these kind of tools is essential not only to understand how these regulatory proteins normally work, but also to understand what happens in a pathological context.