Exploring germline recombination in <scp>Nestin‐Cre</scp> transgenic mice using floxed androgen receptor

McLeod, Victoria M.; Cuic, Brittany; Chiam, Mathew D. F.; Lau, Chew L.; Turner, Bradley J.

doi:10.1002/dvg.23390

Cited by 5 publications

(9 citation statements)

References 22 publications

(23 reference statements)

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“…These findings are most generally of interest to the mouse genetics community. It has long been clear that transgenes carrying Cre under the control of a putatively cell type-specific promoter are often expressed in unexpected tissues ( Song and Palmiter, 2018 ; Stifter and Greter, 2020 ), and here for example our data confirm prior reports of germline activity of Nes Cre and Syn1 Cre drivers ( Luo et al, 2020 ; Betz et al, 1996 ; McLeod et al, 2020 ). Our findings also identify many new examples of off-target Cre activity for a variety of Cre lines, many of which may have been missed before given the relatively small size of the scientific community focused on the epididymis.…”

Section: Discussionsupporting

confidence: 90%

“…Indeed, offspring of the Ai14D; Syn Cre double transgenics exhibited Tomato expression throughout the body, consistent with prior reports of Syn Cre driving germline recombination ( Luo et al, 2020 ). Similarly, the Nestin Cre driver has also been reported to drive germline recombination ( Betz et al, 1996 ; McLeod et al, 2020 ), and we did obtain Tomato-positive offspring of Ai14D; Nes Cre double transgenics. That said, in contrast to the Ai14D; Syn Cre animals we found no detectable labeling of sperm in Ai14D; Nes Cre animals, with Tomato expression in the epididymis largely confined to the epididymal epithelium ( Figure 4—figure supplement 2B ).…”

Section: Resultsmentioning

confidence: 68%

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Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

Rinaldi

Messemer

Desevin

et al. 2023

eLife

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The development of tools to manipulate the mouse genome, including knockout and transgenic technology, has revolutionized our ability to explore gene function in mammals. Moreover, for genes that are expressed in multiple tissues or at multiple stages of development, the use of tissue-specific expression of the Cre recombinase allows gene function to be perturbed in specific cell types and/or at specific times. However, it is well known that putative tissue-specific promoters often drive unanticipated 'off target' expression. In our efforts to explore the biology of the male reproductive tract, we unexpectedly found that expression of Cre in the central nervous system resulted in recombination in the epididymis, a tissue where sperm mature for ~1-2 weeks following the completion of testicular development. Remarkably, we not only observed reporter expression in the epididymis when Cre expression was driven from neuron-specific transgenes, but also when Cre expression in the brain was induced from an AAV vector carrying a Cre expression construct. A surprisingly wide range of Cre drivers - including six different neuronal promoters as well as the adipose-specific Adipoq Cre promoter - exhibited off target recombination in the epididymis, with a subset of drivers also exhibiting unexpected activity in other tissues such as the reproductive accessory glands. Using a combination of parabiosis and serum transfer experiments, we find evidence supporting the hypothesis that Cre may be trafficked from its cell of origin to the epididymis through the circulatory system. Together, our findings should motivate extreme caution when interpreting conditional alleles, and suggest the exciting possibility of inter-tissue RNA or protein trafficking in modulation of reproductive biology.

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Section: Discussionsupporting

confidence: 90%

Section: Resultsmentioning

confidence: 68%

Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

Rinaldi

Messemer

Desevin

et al. 2023

eLife

View full text Add to dashboard Cite

show abstract

“…These findings are most generally of interest to the mouse genetics community. It has long been clear that transgenes carrying Cre under the control of a putatively cell type-specific promoter are often expressed in unexpected tissues [19, 20], and here for example our data confirm prior reports of germline activity of Nes Cre and Syn1 Cre drivers [21–23]. Our findings also identify many new examples of off target Cre activity for a variety of Cre lines, many of which may have been missed before given the relatively small size of the scientific community focused on the epididymis.…”

Section: Discussionsupporting

confidence: 90%

“…Indeed, offspring of the Ai14D; Syn Cre double transgenics exhibited Tomato expression throughout the body, consistent with prior reports of Syn Cre driving germline recombination [21]. Similarly, the Nestin Cre driver has also been reported to drive germline recombination [22,23], and we did obtain Tomato-positive offspring of Ai14D; Nes Cre double transgenics. That said, in contrast to the Ai14D; Syn Cre animals we found no detectable labeling of sperm in Ai14D; Nes Cre animals, with Tomato expression in the epididymis largely confined to the epididymal epithelium (Supplemental Figure S6B).…”

Section: Many-to-many Mapping Between Cre Drivers and Recipient Tissuessupporting

confidence: 90%

Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

Rinaldi

Messemer

Desevin

et al. 2022

Preprint

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The development of tools to manipulate the mouse genome, including knockout and transgenic technology, revolutionized our ability to explore gene function in mammals. Moreover, for genes that are expressed in multiple tissues or at multiple stages of development, the use of tissue-specific expression of the Cre recombinase allows gene function to be perturbed in specific cell types and/or at specific times. However, it is well known that putative tissue-specific promoters often drive unanticipated off target expression. In our efforts to explore the biology of the male reproductive tract, we unexpectedly found that expression of Cre in the central nervous system resulted in recombination in the epididymis, a tissue where sperm mature for ~1-2 weeks following the completion of testicular development. Remarkably, we not only observed reporter expression in the epididymis when Cre expression was driven from neuron-specific transgenes, but also when Cre expression in the brain was induced from an AAV vector carrying a Cre expression construct. A surprisingly wide range of Cre drivers - including six different neuronal promoters as well as the adipose-specific AdipoQ Cre promoter - exhibited off target recombination in the epididymis, with a subset of drivers also exhibiting unexpected activity in other tissues such as the reproductive accessory glands. Finally, using a combination of parabiosis and serum transfer experiments, we find evidence supporting the hypothesis that Cre may be trafficked from its cell of origin to the epididymis through the circulatory system. Together, our findings should motivate extreme caution when interpreting conditional alleles, and suggest the exciting possibility of inter-tissue RNA or protein trafficking in modulation of reproductive biology.

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“…These include damage due to injection, toxicity caused by Cre expression, variable expression, methylationinduced silencing over time, and ectopic expression including in the male and female germlines (Clark, Dinsmore, & Soriano, 2020;Song & Palmiter, 2018). Indeed, Wnt1-Cre2 joins several other transgenic Cre lines recently found to have ectopic male germline activity (de Lange, Halabi, Beyer, & Sigmund, 2008;He et al, 2017;McLeod, Cuic, Chiam, Lau, & Turner, 2020). The ease of transgenesis has historically been viewed as a large enough advantage to overcome the many drawbacks and justify use of the technique.…”

Section: Resultsmentioning

confidence: 99%

The Wnt1‐Cre2 transgene is active in the male germline

Dinsmore

Soriano

2022

Genesis

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The Wnt1-Cre transgenic mouse line is widely used to express the CRE recombinase in neural crest lineages, but it overexpresses WNT1 itself, which can cause undesired phenotypes. To address this, we and others previously developed a Wnt1-Cre2 line based on the same regulatory elements as Wnt1-Cre but without ectopic Wnt1 expression. However, while Wnt1-Cre2 exhibits normal activity when transmitted from female mice, it exhibits unexpected activity in the male germline. The Wnt1-Cre2 transgene was previously mapped to the E2f1 locus. Several genes in this genomic region exhibit significant expression in spermatogonia or spermatocytes, suggesting that local regulatory elements may be driving ectopic transgene expression. The Wnt1-Cre2 line can therefore be used both as a neural crest specific and a general deleter, and care should be taken when setting up genetic crosses.

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Exploring germline recombination in Nestin‐Cre transgenic mice using floxed androgen receptor

Cited by 5 publications

References 22 publications

Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

Evidence for RNA or protein transport from somatic tissues to the male reproductive tract in mouse

The Wnt1‐Cre2 transgene is active in the male germline

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