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Characterization of novel whole body Poldip2 knockout mice generated by [Precisely "flox"ing Poldip2 gene] and crossing with Cre deleter mice

ABSTRACT: Characterization of novel whole body Poldip2 knockout mice generated by [Precisely "flox"ing Poldip2 gene] and crossing with Cre deleter mice

PROVIDER: PRJNA693854 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13499463_1.fastq.gz	Fastqsanger.gz
	SRR13499463_2.fastq.gz	Fastqsanger.gz
	SRR13499464_1.fastq.gz	Fastqsanger.gz
	SRR13499464_2.fastq.gz	Fastqsanger.gz
	SRR13499465_1.fastq.gz	Fastqsanger.gz

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Characterization of novel whole body Poldip2 knockout mice generated by [Precisely "flox"ing Poldip2 gene] and crossing with Cre deleter mice

Project description:Poldip2 is a multifunctional protein whose roles are only partially understood. The previous Poldip2 knockout mouse generated by the gene trap model suffers from two limitations: perinatal lethality in homozygotes and constitutive Poldip2 inactivation. To overcome these limitations, we developed a new conditional floxed Poldip2 mouse. Here, we correctly targeted floxed Poldip2 mice to produce a new constitutive knockout line by crossing with a Cre deleter. In contrast to the gene trap model, many homozygous knockout mice were viable, in spite of having no Poldip2 expression. To characterize the effect of Poldip2 ablation in the vasculature, we performed a RNA-seq study using the carotid arteries of these mice.

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Characterization of novel endothelial-specific and smooth-muscle specific Poldip2 knockout mice

Project description:Poldip2 is a multifunctional protein whose roles are only partially understood. The previous Poldip2 knockout mouse generated by the gene trap model suffers from two limitations: perinatal lethality in homozygotes and constitutive Poldip2 inactivation. To overcome these limitations, we developed a new conditional floxed Poldip2 mouse. Here, we correctly targeted floxed Poldip2 mice to produce an endothelial-specific by crossing it with CDH5 ERT2 Cre mouse. Additionally, we also produced smooth-muscle specific Poldip2 knockout mice by crossing the flox'ed Poldip2 mice with SM22A-Cre mice. The Poldip2_EC-/- and Poldip2_SMC-/-mice were viable and showed remarkably reduced Poldip2 expression. To characterize the effect of Poldip2 ablation in the ECs and in the SMCs, we performed a RNA-seq study using the carotid arteries of these mice.

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Project description:To study the function of AMD-associated gene POLDIP2 in retinal pigment epithelial (RPE) cells, we used CRISPR/Cas to knockout POLDIP2 in the human RPE cell line ARPE19. We then performed RNA-seq to profile the transcriptome of wildtype ARPE19 and POLDIP2 knockout.

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Project description:Maternal inheritance of mitochondrial DNA (mtDNA) is highly conserved in metazoans. While many species eliminate paternal mtDNA during late sperm development to foster maternal inheritance, the regulatory mechanisms governing this process remain elusive. Through a large-scale genetic screen in Drosophila, we identified 47 mutant lines exhibiting substantial retention of mtDNA in mature sperm. We mapped one line to Poldip2, a gene predominantly expressed in the testis. Disruption of Poldip2 led to pronounced mtDNA retention in mature sperm and subsequent paternal transmission to progeny. Further investigation via imaging, biochemical analyses and ChIP assays revealed that POLDIP2 is a mitochondrial matrix protein capable of binding to mtDNA. Moreover, we uncovered that CLPX, a key component of the major mitochondrial protease, binds to POLDIP2 to co-regulate mtDNA elimination in Drosophila spermatids. This study shed light on the mechanisms underlying mtDNA removal during spermatogenesis, underscoring the pivotal role of this process in safeguarding maternal inheritance.

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