The Missing Link: Cre Pigs for Cancer Research

Kalla, Daniela; Flisikowski, Krzysztof; Yang, Kaiyuan; Sangüesa, Laura Beltran; Kurome, Mayuko; Keßler, Barbara; Zakhartchenko, Valeri; Wolf, Eckhard; Lickert, Heiko; Saur, Dieter; Schnieke, Angelika; Flisikowska, Tatiana

doi:10.3389/fonc.2021.755746

Cited by 6 publications

(3 citation statements)

References 35 publications

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“…The use of pigs in this study was approved by the Committee on Animal Health and Care of the local government body of the state of Upper Bavaria in Germany for the wild-type and INS-eGFP 67 German Landrace pigs from the Ludwig Maximilian University of Munich (LMU, Permission No. 55.2-2532.Vet_02-17-136) and the PTF1A-iCre 68 and ROSA-mTmG 69…”

Section: Ethicsmentioning

confidence: 99%

A multimodal cross-species comparison of pancreas development

Lickert,

Yang,

Spitzer

et al. 2024

Preprint

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Human pancreas development remains incompletely understood due to limited sample access constrained by ethical and practical considerations. Here we investigate whether pigs resemble humans in pancreas development more closely than rodents, and as such, offer a valuable alternative large-animal model. As pig pancreas organogenesis is unexplored, we first annotated developmental hallmarks and lineage markers of pancreas differentiation and morphogenesis throughout the 114-day gestation. Building on this detailed roadmap, we further constructed a pig single-cell multiome pancreas atlas capturing temporal resolution across all three trimesters. Cross-species comparisons with human and mouse time-resolved integrated pancreas atlases accentuated that pig closely resembled human in developmental tempo, epigenetic and transcriptional regulation, gene expression patterns and gene regulatory networks (GRNs). Specifically, pig mimicked the dynamics of progenitor status, differentiation trajectories and GRNs governing endocrine fate acquisition in human. In pig multiome GRN, over 40% of transcription factors targeted by NEUROG3, the endocrine master regulator, were confirmed in human stem cell models. Most notably, we uncovered beta-cell heterogeneity arising during embryonic development, owing to endocrine induction in pancreatic progenitors with temporally altered epigenetic and transcriptional identity. Overall, our work lays the foundation for using pigs to model human pancreas biology and provides unprecedented insights into developmental principles and mechanisms across species.

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Section: Ethicsmentioning

confidence: 99%

A multimodal cross-species comparison of pancreas development

Lickert,

Yang,

Spitzer

et al. 2024

Preprint

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“…The recombination of RTs could be induced by mating with Cre-driver animal lines or using a virusbased transduction method to deliver SSRs to targeted organs. Although a variety of mouse strains carrying Cre transgenes have been successfully established [79,80], very few tissue-specific Cre-driver pig lines have been generated by random transgene or CRISPR-mediated knock-in technology [81][82][83]. The main lesson from nearly 30 years of practice of the Cre/ lox system in mice is that each tissue-specific or drug inducible Cre driver line must be carefully evaluated for leakage and efficiency.…”

Section: Compromised Efficiency Of Gene Activationmentioning

confidence: 99%

Genome editing pig models with elements for controllable gene expression

Jin,

Shi,

Liu

et al. 2023

Animal Research and One Health

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Conditional gene regulation systems can control gene expression in predefined tissues or organs at a desired time. Site‐specific recombinase systems and chemically induced gene expression systems are the two most widely used approaches for creating genetically modified (GM) animals with conditional regulation of gene expression. Generation of GM pigs with controllable elements, usually involving multiple gene editing, used to be a major challenge due to a lack of germ line‐competent pluripotent stem cells. With the emergence of artificial endonuclease‐mediated gene editors, a variety of GM pigs with recombinase‐specific recognition elements or chemically induced elements for conditional regulation of gene expression have been generated by the combination of site‐directed knock‐in of somatic cells and somatic cell nuclear transfer technology, allowing conditional deletion of endogenous genes or overexpression of exogenous genes in pigs. These inducible tool pig models will greatly facilitate the production of GM pigs and broaden the applications of transgenic pigs in biomedicine and agriculture fields. In this paper, we review the progress in the construction and application of pigs with controllable elements using gene editing techniques.

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“…Assessing this approach for the porcine ROSA26 locus, we obtained placement efficiencies of nearly 40%. Next, we tested if this method also enabled gene insertions at loci not expressed in the cells used for SCNT and where all traditional gene targeting attempts had previously failed, i.e., placement of Cre-recombinase under the control of an endogenous promoter ( PTF1 ) for tissue-specific pancreatic expression ( Kalla et al, 2021 ). It also allowed the simultaneous addition of (multiple) xeno-relevant transgenes into the intergenic region of a pre-existing transgene locus ( Fischer et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

How genome editing changed the world of large animal research

Fischer,

Schnieke

2023

Front. Genome Ed.

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The first genetically modified large animals were developed in 1985 by microinjection to increase the growth of agricultural livestock such as pigs. Since then, it has been a difficult trail due to the lack of genetic tools. Although methods and technologies were developed quickly for the main experimental mammal, the mouse, e.g., efficient pronuclear microinjection, gene targeting in embryonic stem cells, and omics data, most of it was—and in part still is—lacking when it comes to livestock. Over the next few decades, progress in genetic engineering of large animals was driven less by research for agriculture but more for biomedical applications, such as the production of pharmaceutical proteins in the milk of sheep, goats, or cows, xeno-organ transplantation, and modeling human diseases. Available technologies determined if a desired animal model could be realized, and efficiencies were generally low. Presented here is a short review of how genome editing tools, specifically CRISPR/Cas, have impacted the large animal field in recent years. Although there will be a focus on genome engineering of pigs for biomedical applications, the general principles and experimental approaches also apply to other livestock species or applications.

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The Missing Link: Cre Pigs for Cancer Research

Cited by 6 publications

References 35 publications

A multimodal cross-species comparison of pancreas development

A multimodal cross-species comparison of pancreas development

Genome editing pig models with elements for controllable gene expression

How genome editing changed the world of large animal research

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