Journal: Poultry Science

Article Title: H3K4me2 cooperates with Wnt/TCF7L2 to regulate TDRD1 and promote chicken spermatogonia stem cell formation

doi: 10.1016/j.psj.2023.102552

Figure Lengend Snippet: Effect of H3K4me2 modification on TDRD1. (A) RT-qRCR was used to detect TDRD1 expression after OE-TCF7L2 or sh-TCF7L2. * P < 0.05, ** P < 0.01 (B) Expression of TDRD1 in the process of inducing ESCs to SSCs in vitro was detected by RT-qPCR. (C) Expression of TDRD1 in SSCs isolated from testis after interference with MLL2 and LSD1 was detected by RT-qRCR on day 18.5. (D) Western blotting was used to detect the expression of related proteins in vivo after interference with MLL2 and LSD1. (E) Grayscale analysis of western blot image.

Article Snippet: sgRNAs were designed every 200 bp according to the TDRD1 promoter enrichment region on the CRISPOR website ( http://crispor.tefor.net/crispor.py ) , add the cleavage site of BbsI enzyme and the protective base, annealing, and amplification. pX330a dCas9 LSD1 vector was purchased from Addgene.

Techniques: Modification, Expressing, In Vitro, Quantitative RT-PCR, Isolation, Western Blot, In Vivo

Journal: Poultry Science

Article Title: H3K4me2 cooperates with Wnt/TCF7L2 to regulate TDRD1 and promote chicken spermatogonia stem cell formation

doi: 10.1016/j.psj.2023.102552

Figure Lengend Snippet: Construction of dCas9-LSD1 vector targeting P1/P2/P3/P4 of TDRD1 promoter. (A) Schematic diagram of H3K4me2 and TCF7L2 in the TDRD1 promoter region. (B) Structure diagram of pX330a dCas9. (C) Detection of H3K4me2 levels in the TDRD1 promoter region by ChIP-qPCR. (D) ChIP-qPCR detection of different dCas9-LSD1 targeting levels of H3K4me2 in the P1/P2/P3/P4 regions of the TDRD1 promoter. (E, F) Effect of H3K4me2 levels on TDRD1 initiating activity detected by dual luciferase reporter assays.

Article Snippet: sgRNAs were designed every 200 bp according to the TDRD1 promoter enrichment region on the CRISPOR website ( http://crispor.tefor.net/crispor.py ) , add the cleavage site of BbsI enzyme and the protective base, annealing, and amplification. pX330a dCas9 LSD1 vector was purchased from Addgene.

Techniques: Plasmid Preparation, Activity Assay, Luciferase

Journal: Genome Medicine

Article Title: Transcriptome-wide association study revealed novel causal genes of renal-biopsy proven diabetic nephropathy

doi: 10.1186/s13073-025-01590-x

Figure Lengend Snippet: The function exploration of genetic variants around GYS1 . Expression quantitative trait loci (eQTL) box plot. Box plot displaying the effect of the genotypes on gene expression levels in kidney tissues. Genotypic effects of ( A ) SNPs rs111518556 and ( B ) rs116573491 on expression of GYS1. The y axis shows the gene expression levels, and the x axis shows the genotypes of SNPs. The box plot displays the first and third quartiles (top and bottom of the boxes), the median (band inside the boxes), and the lowest and highest point within 1.5 times the interquartile range of the lower and higher quartiles. ( C ) The functional annotation for SNPs located near GYS1 in ENCODE and Roadmap. CRIPSR-dCas9-VP64-mediated transcriptional activation ( D ) in HEK293T cells on rs11518556 ( E ) and rs116573491 ( F )

Article Snippet: Human embryonic kidney (HEK293T) cells were purchased from the American Type Culture Collection (ATCC), HEK293T was grown in Dulbecco’s modified Eagle’s Medium (DMEM) (VivaCell, C3114-0500) supplemented with 10% FBS + 1% penicillin/streptomycin. pX330a dCas9-VP64 was a gift from Tatjana Sauka-Spengler (Addgene plasmid # 92,363), sgRNAs guide sequences were cloned into pSpCas9 (BB)−2A-Neo which was a gift from Ken-Ichi Takemaru (Addgene plasmid # 127,762).

Techniques: Expressing, Gene Expression, Functional Assay, Activation Assay