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首页> 外文期刊>PLoS One >A novel mouse line with epididymal initial segment-specific expression of Cre recombinase driven by the endogenous Lcn9 promoter
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A novel mouse line with epididymal initial segment-specific expression of Cre recombinase driven by the endogenous Lcn9 promoter

机译:一种新的小鼠线,具有由内源性LCN9启动子驱动的CRE重组酶的附睾初始段特异性表达的新型鼠标系列

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摘要

Spermatozoa released from testes undergo a maturation process and acquire the capacity to fertilize ova through epididymal transit. The epididymis is divided into four regions, each with unique morphology, gene profile, luminal microenvironment and distinct function. To study the functions of relevant genes in the epididymal initial segment (IS), a novel IS-specific mouse model, Lcn9-Cre knock-in (KI) mouse line was generated via CRISPR/Cas9 technology. The TAG stop codon was replaced by a 2A-NLS-Cre cassette, resulting in the co-expression of Lcn9 and Cre recombinase. IS-specific Cre expression was first observed from postnatal day 17. Using the Rosa26 tdTomato reporter mice, the Cre-mediated DNA recombination was detected exclusively in principal cells. The epididymal IS-specific Cre activity in vivo was further confirmed using Lcn9-Cre mice crossed with a mouse strain carrying Tsc1 floxed alleles ( Tsc1 flox/+ ). Cre expression did not affect either normal development or male fecundity. Different from any epididymis-specific Cre mice reported previously, the novel Lcn9-Cre mouse line can be used to introduce entire IS-specific conditional gene editing for gene functional study.
机译:从睾丸中释放的精子经历成熟过程,并通过附睾转运获得卵子的能力。附睾分为四个区域,每个地区具有独特的形态,基因曲线,腔微环境和不同的功能。为了研究附睾初始段(IS)中的相关基因的功能,通过CRISPR / CAS9技术产生一种新的特异性小鼠模型,LCN9-CRE敲入(KI)小鼠线。标签停止密码子被2A-NLS-CRE盒代替,导致LCN9和CRE重组酶的共表达。首先从后期第17天开始观察到特异性CRE表达。使用ROSA26 TDTOMATO报告机构,仅在主要细胞中仅检测CRE介导的DNA重组。使用携带TSC1氟化等位基因(TSC1 FLOX / +)的小鼠菌株交叉进一步证实了体内的eVIACIDMAL特异性CRE活性。 CRE表达没有影响正常的发展或男性繁殖力。不同于先前报道的任何附睾特异性CRE小鼠,新型LCN9-CRE小鼠线可用于引入基因功能研究的整个特异性条件基因编辑。

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