Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome

Sakata, Kazuya; Araki, Kimi; Nakano, Hiroyasu; Nishina, Takashi; Komazawa-Sakon, Sachiko; Murai, S.; Lee, Grace E.; Hashimoto, Daisuke; Suzuki, Chigure; Uchiyama, Yasuo; Notohara, Kenji; Gukovskaya, Anna S.; Gukovsky, Ilya; Yamamura, Ken Ichi; Baba, Hideo; Ohmuraya, Masaki

doi:10.1038/srep37200

Cited by 13 publications

(5 citation statements)

References 32 publications

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“…In the histopathologic examination, the Ctsb ΔPan ;Ctsd ΔPan mouse pancreas showed CP, which progressed with time. At least four animal models of CP with impaired autophagy have been previously reported: mice with pancreas-specific disruption of Atg5 that developed a form of CP 4 , mice with pancreas-specific ablation of IκB kinase α 5 , mice lacking Spink3 with a mosaic pattern of SPINK1 expression 6 , and LAMP-2-deficient mice 7 . The histological findings of CP, such as inflammation, acinar-to-ductal metaplasia and acinar-cell hypertrophy, have been confirmed in all these models.…”

Section: Discussionmentioning

confidence: 99%

Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis

Iwama

Mehanna

Imasaka

et al. 2021

Sci Rep

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The major lysosomal proteases, Cathepsin B (CTSB), Cathepsin D (CTSD) and Cathepsin L (CTSL), are implicated in autophagic activity. To investigate the role of each cathepsin in the exocrine pancreas, we generated mice in which the pancreas was specifically deficient in Ctsb, Ctsd and Ctsl. Each of these gene knockout (KO) and Ctsb;Ctsl and Ctsd;Ctsl double-knockout (DKO) mice were almost normal. However, we found cytoplasmic degeneration in the pancreatic acinar cells of Ctsb;Ctsd DKO mice, similar to autophagy related 5 (Atg5) KO mice. LC3 and p62 (autophagy markers) showed remarkable accumulation and the numbers of autophagosomes and autolysosomes were increased in the pancreatic acinar cells of Ctsb;Ctsd DKO mice. Moreover, these Ctsb;Ctsd DKO mice also developed chronic pancreatitis (CP). Thus, we conclude that both Ctsb and Ctsd deficiency caused impaired autophagy in the pancreatic acinar cells, and induced CP in mice.

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Section: Discussionmentioning

confidence: 99%

Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis

Iwama

Mehanna

Imasaka

et al. 2021

Sci Rep

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“…Despite the recent progress, detailed analyses of organelle disorders in exocrine pancreas have just begun and there is much to be learned about their role in pancreatitis and the underlying mechanisms. 3,4,[134][135][136][137][138][139][140][141][142][143] One important area is the interrelations between different types of organelles in acinar cells and how these are altered by pancreatitis. A key question is whether there is a critical pathologic defect common for all types of pancreatitis and leading to failure of the whole organellar network, or whether organelle dysfunction is mediated through different mechanisms and more than one organelle disorder should be restored (for example, normalizing both autophagy and mitochondrial function at the same time) to re-establish the organellar network homeostasis.…”

Section: Future Directionsmentioning

confidence: 99%

Recent Insights Into the Pathogenic Mechanism of Pancreatitis

Gukovskaya¹,

Gorelick²,

Groblewski

et al. 2019

Pancreas

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Acute pancreatitis (AP) is a potentially lethal inflammatory disease that lacks specific therapy. Damaged pancreatic acinar cells are believed the site of AP initiation. The primary function of these cells is the synthesis, storage, and export of digestive enzymes. Beginning in the endoplasmic reticulum and ending with secretion of proteins stored in zymogen granules, distinct pancreatic organelles use ATP produced by mitochondria to move and modify nascent proteins through sequential vesicular compartments. Compartment-specific accessory proteins concentrate cargo, promote vesicular budding, targeting and fusion. The autophagy-lysosomal-endosomal pathways maintain acinar cell homeostasis by removing damaged/dysfunctional organelles and recycling cell constituents for substrate and energy. Here, we discuss studies in experimental and genetic AP models, primarily from our groups, which show that acinar cell injury is mediated by distinct mechanisms of organelle dysfunction involved in protein synthesis and trafficking, secretion, energy generation, and autophagy. These early AP events (often first manifest by abnormal cytosolic Ca 2+ signaling) in the acinar cell trigger the inflammatory and cell death

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“…However, using cell-specific promoters to drive cre, as in this study, should reduce or eliminate this challenge. The unique opportunity is that when the cre is brought through either the maternal or paternal parent, X-chromosome inactivation mosaic heterozygous females are obtained; mosaicism can be a useful analytical tool, as exemplified by this study of expression and others (Sakata et al 2016; Renthal et al 2018).…”

Section: Discussionmentioning

confidence: 99%

Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye and Brain, Including Seventeen Carrying a New Inducible-First Constitutive-Ready Allele

et al. 2019

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To understand gene function, the cre/loxP conditional system is the most powerful available for temporal and spatial control of expression in mouse. However, the research community requires more cre recombinase expressing transgenic mouse strains (cre-drivers) that restrict expression to specific cell types. To address these problems, a high-throughput method for large-scale production that produces high-quality results is necessary. Further, endogenous promoters need to be chosen that drive cell type specific expression, or we need to further focus the expression by manipulating the promoter. Here we test the suitability of using knock-ins at the docking site 5′ of Hprt for rapid development of numerous cre-driver strains focused on expression in adulthood, using an improved cre tamoxifen inducible allele (icre/ERT2), and testing a novel inducible-first, constitutive-ready allele (icre/f3/ERT2/f3). In addition, we test two types of promoters either to capture an endogenous expression pattern (MaxiPromoters), or to restrict expression further using minimal promoter element(s) designed for expression in restricted cell types (MiniPromoters). We provide new cre-driver mouse strains with applicability for brain and eye research. In addition, we demonstrate the feasibility and applicability of using the locus 5′ of Hprt for the rapid generation of substantial numbers of cre-driver strains. We also provide a new inducible-first constitutive-ready allele to further speed cre-driver generation. Finally, all these strains are available to the research community through The Jackson Laboratory.

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Novel method to rescue a lethal phenotype through integration of target gene onto the X-chromosome

Cited by 13 publications

References 32 publications

Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis

Cathepsin B and D deficiency in the mouse pancreas induces impaired autophagy and chronic pancreatitis

Recent Insights Into the Pathogenic Mechanism of Pancreatitis

Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye and Brain, Including Seventeen Carrying a New Inducible-First Constitutive-Ready Allele

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