A novel F‐box domain containing cyclin F like gene is required for maintaining the genome stability and survival of chicken primordial germ cells

Abstract

The stability and survival of germ cells are controlled by the germline‐specific genes, however, such genes are less known in the avian species. Using a microarray‐based the National Center for Biotechnology Information Gene Expression Omnibus dataset, we found an unigene (Gga.9721) that upregulated in the chicken primordial germ cells (PGCs). The unigene showed 97% identities with an uncharacterized chicken cyclin F like gene. The predicted chicken cyclin F like gene was further characterized through expression and regulation in the chicken PGCs. The sequence analysis revealed that the gene shows identities with cyclin F gene and contains an F‐box domain. The expression of chicken cyclin F like was detected specifically in the gonads, PGCs, and germline cells. The knockdown of cyclin F like gene resulted in DNA damage and apoptosis in the PGCs. The genes related to stemness and germness were downregulated, whereas, genes related to apoptosis and DNA damage response were increased in the PGCs after the knockdown of chicken cyclin F like. We further observed that the Nanog homeobox controlled the transcriptional activity of chicken cyclin F like gene in PGCs. Collectively, the chicken cyclin F like gene, which is not reported in any other species, is required for maintaining the genome stability of germ cells.

Chicken blastoderms and primordial germ cells possess a higher expression of DNA repair genes and lower expression of apoptosis genes to preserve their genome stability

“…Several phosphoinositide 3-kinase-related protein kinases, such as ataxia teleangiectasia mutated (ATM), ATM and Rad3-related (ATR), or DNA-dependent protein kinase (DNA-PK), are considered as the major physiological mediator of H2AX phosphorylation 22 , 52 . Analysis of phosphorylated H2AX (γ-H2A.X) expression is widely used to detect the genotoxic effect in cells exposed to various toxic substances and in cells where genome stability-related genes are affected 52 – 54 . According to our results of the TUNEL assay, DNA damage was successfully induced in CEFs, blastoderm cells, and PGCs after treatment with H 2 O 2 .…”

Robust Three-Dimensional (3D) Expansion of Bovine Intestinal Organoids: An In Vitro Model as a Potential Alternative to an In Vivo System

“…Total RNA for prepared samples including intestinal organoids was isolated using TRIzol reagent (Life Technologies, Carlsbad, CA, USA), as described previously [25,26]. RNA quality was assessed by an Agilent 2100 bioanalyser using an RNA 6000 nano chip (Agilent Technologies, Amstelveen, The Netherlands), and RNA quantification was performed using an ND 2000 spectrophotometer (Thermo Inc., Wilmington, DE, USA).…”

Research Note: Development of a chicken experimental model platform for induced pluripotent stem cells by using CRISPR/Cas9-mediated NANOG knock-in reporter DF1 cells

“…Total RNA from prepared samples, including Eyal-Giladi and Kochav ( EGK ) stage X embryos and chicken induced pluripotent stem cells ( ciPSC s), was isolated using TRIzol reagent (Life Technologies, Carlsbad, CA) as described previously ( Lee et al., 2020 ), and quantitative RT‒PCR was performed to assess the expression of pluripotent markers, as summarized in Table 1 . The PCR mixture was prepared by adding 2 μL of 10 pmol of each forward and reverse primer, 7 μL of nuclease-free water, 10 μL of SYBR Green qPCR Mater Mix, and 1 μL of cDNA to a final volume of 20 μL.…”