A novel conditional ZsGreen-expressing transgenic reporter rat strain for validating Cre recombinase expression

Bryda, Elizabeth C.; Men, Hongsheng; Davis, Daniel R.; Bock, Anagha S.; Shaw, Mary L.; Chesney, Kari L.; Hankins, Miriam A.

doi:10.1038/s41598-019-49783-w

Cited by 16 publications

(11 citation statements)

References 17 publications

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“…The CRISPR crRNA/tracrRNA target site is located close to the Prl7b1 ATG start codon. (C) The reporter rat consists of a ubiquitously active CAG promoter (CAG), a LoxP-3x Stop-LoxP cassette (LSL) and ZsGreen ( Bryda et al, 2019 ). The ZsGreen protein is expressed only where Prl7b1-iCre is expressed.…”

Section: Resultsmentioning

confidence: 99%

Conditionally mutant animal model for investigating the invasive trophoblast cell lineage

Iqbal,

Dominguez,

Nixon

et al. 2024

Development

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Placental development involves coordinated expansion and differentiation of trophoblast cell lineages possessing specialized functions. Among the differentiated trophoblast cell lineages are invasive trophoblast cells, which exit the placenta and invade into the uterus where they restructure the uterine parenchyma and facilitate remodeling of uterine spiral arteries. The rat exhibits deep intrauterine trophoblast cell invasion, a feature shared with human placentation, and is also amenable to gene manipulation using genome editing techniques. In this investigation, we generated a conditional rat model targeting the invasive trophoblast cell lineage. Prolactin family 7, subfamily b, member 1 (Prl7b1) is uniquely and abundantly expressed in the rat invasive trophoblast cell lineage. Disruption of Prl7b1 did not adversely affect placental development. We demonstrated that the Prl7b1 locus could be effectively used to drive the expression of Cre recombinase in invasive trophoblast cells. Our rat model represents a new tool for investigating candidate genes contributing to the regulation of invasive trophoblast cells and their contributions to trophoblast-guided uterine spiral artery remodeling.

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Section: Resultsmentioning

confidence: 99%

Conditionally mutant animal model for investigating the invasive trophoblast cell lineage

Iqbal,

Dominguez,

Nixon

et al. 2024

Development

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“…Detection probe for the ZsGreen1 transgene: 5’-/56-FAM/TT CAT CCA G/ZEN/C ACA AGC TGA C/3IABkFQ/-3’ (Bryda et al ., 2019) (synthesized by Integrated DNA Technologies, Inc.)…”

Section: Methodsmentioning

confidence: 99%

“…Primers for the ZsGreen1 transgene: 5’-GTG TAC AAG GCC AAG TCC GT-3’ and 5’-CCA CTT CTG GTT CTT GGC GT-3’ (Bryda et al ., 2019) (synthesized by Eurofin Genomics K.K. )…”

Section: Methodsmentioning

confidence: 99%

Purification Method for Adeno-Associated Viral Vector Serotype 9 Using Ceramic Hydroxyapatite Chromatography and Its Qualification

Kurosawa,

Tsunekawa,

Wada

et al. 2024

Preprint

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Adeno-associated virus (AAV) vectors can efficiently transduce exogenous genes into various tissues in vivo. Owing to their convenience, high efficiency, long-term stable gene expression, and minimal side effects, AAV vectors have become one of the gold standards for investigating gene functions in vivo, especially in non-clinical studies. However, challenges persist in efficiently preparing a substantial quantity of high-quality AAV vectors that comprise only genome-packaged full particles. Commercial AAV vectors are typically associated with high costs. Further, in-laboratory production is hindered by the lack of specific laboratory equipment, such as ultracentrifuges. Therefore, a simple, quick, and scalable preparation method for AAV vectors is needed for proof-of-concept experiments. Herein, we present an optimized method for producing and purifying high-quality AAV serotype 9 (AAV9) vectors using standard laboratory equipment and chromatography. Using ceramic hydroxyapatite as a mixed-mode chromatography medium can markedly increase the quality of purified AAV vectors. Basic protocols and optional methods for evaluating purified AAV vectors are also described.

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“…By crossbreeding fluorophore loxP "floxed" mice [67][68][69] or rats [69][70][71][72] with neuronal cell-typespecific Cre-expressing mice [73][74][75] or rats [76][77][78], we can label cell-type-specific synapses for MSbased neuroproteomics analyses and other biochemical assays (Figure 2A). In addition, Credependent viruses can be applied to a specific brain region of neuron-specific Cre animals to label a subcellular compartment of synapses [66].…”

Section: Transgenic Animalsmentioning

confidence: 99%

Cell-type-specific neuroproteomics of synapses

Yim¹,

Nestler²

2023

Preprint

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In the last two decades, our knowledge of synaptic proteomes and their relationship to normal brain function and neuropsychiatric disorders has been expanding rapidly through the use of more powerful neuroproteomic approaches. However, mass spectrometry (MS) based neuroproteomic studies of synapses still need cell-type, spatial, and temporal proteome information. With the advancement of sample preparation and MS techniques, we have just begun to identify and understand proteomes within a given cell type, subcellular compartment, and cell-type-specific synapse. Here, we review the progress and limitations of MS-based neuroproteomics of synapses in the mammalian CNS and highlight the recent applications of these approaches in studying neuropsychiatric disorders such as major depressive disorder and substance use disorders. Combining neuroproteomic findings with other omics studies can generate an in-depth, comprehensive map of synaptic proteomes and possibly identify new therapeutic targets and biomarkers of several central nervous system disorders.

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A novel conditional ZsGreen-expressing transgenic reporter rat strain for validating Cre recombinase expression

Cited by 16 publications

References 17 publications

Conditionally mutant animal model for investigating the invasive trophoblast cell lineage

Conditionally mutant animal model for investigating the invasive trophoblast cell lineage

Purification Method for Adeno-Associated Viral Vector Serotype 9 Using Ceramic Hydroxyapatite Chromatography and Its Qualification

Cell-type-specific neuroproteomics of synapses

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