Simultaneous deletion of floxed genes mediated by CaMKIIα-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Goα

Choi, Chan-Il; Yoon, Sang-Phil; Choi, Jung-Mi; Kim, Sung‐Soo; Lee, Young‐Don; Birnbaumer, Lutz; Suh‐Kim, Haeyoung

doi:10.1038/emm.2014.14

Cited by 27 publications

(23 citation statements)

References 33 publications

(45 reference statements)

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“…First is that Foxn1, and constructs regulated by this locus, joins an expanding list of genes expressed in tissues where they were not generally expected, often including the male germline, see [9,10–12] for examples. Expression of Foxn1[Cre] in the male germline can be a useful attribute, since it facilitates generation of knockout sperm in males heterozygous for genes that are lethal when homozygous.…”

Section: Discussionmentioning

confidence: 99%

Foxn1[Cre] Expression in the Male Germline

et al. 2016

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Foxn1 (forkhead box N1), also known as the nude gene or winged-helix nude (Whn), is a forkhead transcription factor thought to be restricted to keratinocytes in the skin and thymus. Consistent with this tissue distribution, spontaneous or targeted mutation of Foxn1 results in the absence of both hair and a thymus. Genetic manipulation of the Foxn1 locus thus represents a powerful tool for tissue specific gene control in the skin and thymus, and tools such as Cre recombinase under control of the Foxn1 locus are widely used for this purpose. Unexpectedly, we show that Foxn1[Cre] exhibits unexpected activity in male germ cells, resulting in ubiquitous targeting of loxP-flanked alleles in all tissues in offspring from Foxn1[Cre] expressing male mice. Inheritance of recombined loxP alleles occurs independently of Cre inheritance (i.e., offspring lacking Cre nonetheless exhibit recombined alleles), suggesting that Foxn1[Cre] induced recombination in male germ cells must occur prior to meiosis in diploid germ cells. Together with previously published data, our results show that Foxn1, and alleles under its control, are expressed in the pre-meiotic male germline, revealing a new tool for germline targeting of genes, and raising important concerns for gender selection when using Foxn1 regulatory elements.

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Section: Discussionmentioning

confidence: 99%

Foxn1[Cre] Expression in the Male Germline

et al. 2016

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“…Given that the Camk2a-cre lines they used can induce a simultaneous deletion of floxed Htt gene in the brain and male germ cells (Choi et al, 2014), the possibility that the sterility they observed could be due to an impaired hypothalamic-pituitary-testis axis caused by the Cre-mediated recombination in the brain could not be excluded. In this study, we generated a CKO mouse that had conditional knockout of Htt specifically in the male germ cells.…”

Section: Discussionmentioning

confidence: 99%

“…These results hint that Htt might play an essential role during spermatogenesis in testis. However, considering that the Camk2a-cre lines they employed can induce a simultaneous deletion of floxed Htt gene in the brain and male germ cells (Choi et al, 2014), it is necessary to generate a germ-cell-conditional knockout mouse (CKO) model to directly confirm the possibility.…”

Section: Introductionmentioning

confidence: 99%

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

Yan¹,

Zhang²,

Liu³

et al. 2016

Development

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Human Huntingtin (HTT), a Huntington's disease gene, is highly expressed in the mammalian brain and testis. Simultaneous knockout of mouse Huntingtin (Htt) in brain and testis impairs male fertility, providing evidence for a link between Htt and spermatogenesis; however, the underlying mechanism remains unclear. To understand better the function of Htt in spermatogenesis, we restricted the genetic deletion specifically to the germ cells using the Cre/loxP sitespecific recombination strategy and found that the resulting mice manifested smaller testes, azoospermia and complete male infertility. Meiotic chromosome spread experiments showed that the process of meiosis was normal in the absence of Htt. Notably, we found that Httdeficient round spermatids did not progress beyond step 3 during the post-meiotic phase, when round spermatids differentiate into mature spermatozoa. Using an iTRAQ-based quantitative proteomic assay, we found that knockout of Htt significantly altered the testis protein profile. The differentially expressed proteins exhibited a remarkable enrichment for proteins involved in translation regulation and DNA packaging, suggesting that Htt might play a role in spermatogenesis by regulating translation and DNA packaging in the testis.

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“…Expression of Cre transgene in the testis was also reported in other Camk2α-Cre transgenic mouse lines and the recombination efficiency in the testis differed depending on floxed genes. 24…”

Section: Generation Of Constitutive and Neuronspecific Crls1 Knockomentioning

confidence: 99%

Cardiolipin is essential for early embryonic viability and mitochondrial integrity of neurons in mammals

et al. 2019

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Cardiolipin (CL) is a hallmark phospholipid of mitochondria and plays a significant role in maintaining the mitochondrial structure and functions. Despite the physiological importance of CL, mutant organisms, yeast, Arabidopsis, C elegans, and Drosophila, which lack CL synthase (Crls1) gene and consequently are deprived of CL, are viable. Here we report conditional Crls1-deficient mice using targeted insertion of loxP sequences flanking the functional domain of CRLS1 enzyme.Homozygous null mutant mice exhibited early embryonic lethality at the peri-implantation stage. We generated neuron-specific Crls1 knockout (cKO) mice by crossing with Camk2α-Cre mice. Neuronal loss and gliosis were gradually manifested in the forebrains, where CL levels were significantly decreased. In the surviving neurons, malformed mitochondria with bubble-like or onion-like inner membrane structures were observed. We showed decreased supercomplex assembly and reduced enzymatic activities of electron transport chain complexes in the forebrain of cKO mice, resulting in affected mitochondrial calcium dynamics, a slower rate of Ca 2+ uptake and a smaller calcium retention capacity. These observations clearly demonstrate indispensable roles of CL as well as of Crls1 gene in mammals.

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Simultaneous deletion of floxed genes mediated by CaMKIIα-Cre in the brain and in male germ cells: application to conditional and conventional disruption of Goα

Cited by 27 publications

References 33 publications

Foxn1[Cre] Expression in the Male Germline

Foxn1[Cre] Expression in the Male Germline

Germline deletion of huntingtin causes male infertility and arrested spermiogenesis in mice

Cardiolipin is essential for early embryonic viability and mitochondrial integrity of neurons in mammals

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