Using TARGATT™ Technology to Generate Site-Specific Transgenic Mice

Chen-Tsai, Ruby Yanru

doi:10.1007/978-1-4939-8831-0_4

Cited by 5 publications

(4 citation statements)

References 23 publications

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“…To create the KO + hTRIM28 mice, a construct encoding HA-tagged hTRIM28 (HA-hTRIM28) downstream of a LoxP-Stop-LoxP (LSL) cassette was knocked into the H11 locus of TARGATT TM C57BL6 mice ( 54 ) (Fig. S9A-B).…”

Section: Resultsmentioning

confidence: 99%

“…The HA-hTRIM28 construct was then ligated into the multiple cloning site (MCS) of TARGATT TM 6.1 (CAG – L4SL – MCS – PolyA) vector which contains a LoxP-StopP-LoxP (LSL) cassette and an attB integration sequence (Applied StemCell, #AST-3050). Embryos from TARGATT TM H11 mice (Applied StemCell) which contained an attP integration site in the H11 locus were injected with ΦC31 integrase plus TARGATT TM 6.1-LSL-HA-hTRIM28 ( 54 ). The offspring with successful integration were crossed with the Pax7 -iCre +/- : Trim28 flox/flox mice to generate (KO + hTRIM28) mice with the genotype Pax7 -iCre +/- : Trim28 flox/flox : LSL-HA-hTRIM28 +/+ .…”

Section: Methodsmentioning

confidence: 99%

“…Embryos from TARGATT TM H11 mice (Applied StemCell) which contained an attP integration site in the H11 locus were injected with ΦC31 integrase plus TARGATT TM 6.1-LSL-HA-hTRIM28 (54). The offspring with successful integration were crossed with the Pax7-iCre +/-: Trim28 flox/flox mice to generate (KO + hTRIM28) mice with the genotype Pax7-iCre +/-: Trim28 flox/flox : LSL-HA-hTRIM28 +/+ .…”

Section: Animalsmentioning

confidence: 99%

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The role of satellite cell-derived TRIM28 in mechanical load- and injury-induced myogenesis

Lin,

Hibbert,

Lemens

et al. 2023

Preprint

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Satellite cells are skeletal muscle stem cells that contribute to postnatal muscle growth, and they endow skeletal muscle with the ability to regenerate after a severe injury. Here we discovered that this myogenic potential of satellite cells requires a protein called tripartite motif-containing 28 (TRIM28). Unexpectedly, multiple lines of both in vitro and in vivo evidence revealed that the myogenic function of TRIM28 is not dependent on changes in the phosphorylation of its serine 473 residue. Moreover, the functions of TRIM28 were not mediated through the regulation of satellite cell proliferation or differentiation. Instead, our findings indicate that TRIM28 regulates the ability of satellite cells to progress through the process of fusion. Specifically, we discovered that TRIM28 controls the expression of a fusogenic protein called myomixer and concomitant fusion pore formation. Collectively, the outcomes of this study expose the framework of a novel regulatory pathway that is essential for myogenesis.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Animalsmentioning

confidence: 99%

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The role of satellite cell-derived TRIM28 in mechanical load- and injury-induced myogenesis

Lin,

Hibbert,

Lemens

et al. 2023

Preprint

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“…SELECTIV mice were generated at the Stanford Transgenic, Knockout, and Tumor Model Center using Integrase Mediated Transgenesis 74 and using the construct described above and PhiC31 integrase. The construct was inserted into the H11 locus using C57BL/6 mice with three attP sites previously knocked-in to the H11 locus.…”

Section: Methodsmentioning

confidence: 99%

Hardwiring tissue-specific AAV transduction in mice through engineered receptor expression

et al. 2023

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The development of transgenic mouse models that express genes of interest in specific cell types has transformed our understanding of basic biology and disease. However, generating these models is time- and resource-intensive. Here we describe a model system, SELective Expression and Controlled Transduction In Vivo (SELECTIV), that enables efficient and specific expression of transgenes by coupling adeno-associated virus (AAV) vectors with Cre-inducible overexpression of the multi-serotype AAV receptor, AAVR. We demonstrate that transgenic AAVR overexpression greatly increases the efficiency of transduction of many diverse cell types, including muscle stem cells, which are normally refractory to AAV transduction. Superior specificity is achieved by combining Cre-mediated AAVR overexpression with whole-body knockout of endogenous Aavr, which is demonstrated in heart cardiomyocytes, liver hepatocytes and cholinergic neurons. The enhanced efficacy and exquisite specificity of SELECTIV has broad utility in development of new mouse model systems and expands the use of AAV for gene delivery in vivo.

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Alterations in the spatiotemporal expression of the chemokine receptor CXCR4 in endothelial cells cause failure of hierarchical vascular branching

Liu

Burns

et al. 2021

Developmental Biology

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Using TARGATT™ Technology to Generate Site-Specific Transgenic Mice

Cited by 5 publications

References 23 publications

The role of satellite cell-derived TRIM28 in mechanical load- and injury-induced myogenesis

The role of satellite cell-derived TRIM28 in mechanical load- and injury-induced myogenesis

Hardwiring tissue-specific AAV transduction in mice through engineered receptor expression

Alterations in the spatiotemporal expression of the chemokine receptor CXCR4 in endothelial cells cause failure of hierarchical vascular branching

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