The History of Transgenesis

Saunders, Thomas L.

doi:10.1007/978-1-4939-9837-1_1

Cited by 13 publications

(8 citation statements)

References 175 publications

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“…Historically, the transgenic mouse field operated under the convention that a minimum of three independent genetic events yielding the same outcome would be required to establish that the observed phenotypic outcome was caused by the genetic manipulation 19 , as opposed to some random parameter, which would not be expected to occur consistently among multiple individuals. However, in cases of incomplete penetrance or phenotype variability (also termed variable expressivity), larger sample numbers are necessary to document the phenotypic spectrum with convincing robustness.…”

Section: Discussionmentioning

confidence: 99%

Broad spectrum of CRISPR-induced edits in an embryonic lethal gene

Fuselier

Kappen

2021

Sci Rep

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Mendelian genetics poses practical limitations on the number of mutant genes that can be investigated simultaneously for their roles in embryonic development in the mouse. While CRISPR-based gene editing of multiple genes at once offers an attractive alternative strategy, subsequent breeding or establishment of permanent mouse lines will rapidly segregate the different mutant loci again. Direct phenotypic analysis of genomic edits in an embryonic lethal gene in F0 generation mice, or F0 mouse embryos, circumvents the need for breeding or establishment of mutant mouse lines. In the course of genotyping a large cohort of F0 CRISPants, where the embryonic lethal gene T/brachyury was targeted, we noted the presence of multiple CRISPR-induced modifications in individual embryos. Using long-read single-molecule Nanopore sequencing, we identified a wide variety of deletions, ranging up to 3 kb, that would not have been detected or scored as wildtype with commonly used genotyping methods that rely on subcloning and short-read or Sanger sequencing. Long-read sequencing results were crucial for accurate genotype–phenotype correlation in our F0 CRISPants. We thus demonstrate feasibility of screening manipulated F0 embryos for mid-gestation phenotypic consequences of CRISPR-induced mutations without requiring derivation of permanent mouse lines.

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Section: Discussionmentioning

confidence: 99%

Broad spectrum of CRISPR-induced edits in an embryonic lethal gene

Fuselier

Kappen

2021

Sci Rep

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“…Similar to ZFN, transcription activator‐like effector nucleases (TALENs) that rely on the Fok1 endonuclease were first used to generate genetically modified embryos in 2011 24 . Although ZFN and TALEN have been widely used to generate genetically modified embryos, they have now been replaced by clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR associated (Cas9) because of their simplicity and flexibility in targeting 25 . Recently, there have been a number of comparative studies in the mammalian embryo field, 26 which have used the CRISPR/Cas9 technology to accelerate research 27,28 .…”

Section: Introductionmentioning

confidence: 99%

“… 24 Although ZFN and TALEN have been widely used to generate genetically modified embryos, they have now been replaced by clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR associated (Cas9) because of their simplicity and flexibility in targeting. 25 Recently, there have been a number of comparative studies in the mammalian embryo field, 26 which have used the CRISPR/Cas9 technology to accelerate research. 27 , 28 However, since only a limited number of species have been studied, we tried to determine the pluripotent network in the pig embryo, which is recognized as a model animal.…”

Section: Introductionmentioning

confidence: 99%

SOX2 plays a crucial role in cell proliferation and lineage segregation during porcine pre‐implantation embryo development

Lee

Choi

et al. 2021

Cell Proliferation

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Objectives Gene regulation in early embryos has been widely studied for a long time because lineage segregation gives rise to the formation of a pluripotent cell population, known as the inner cell mass (ICM), during pre‐implantation embryo development. The extraordinarily longer pre‐implantation embryo development in pigs leads to the distinct features of the pluripotency network compared with mice and humans. For these reasons, a comparative study using pre‐implantation pig embryos would provide new insights into the mammalian pluripotency network and help to understand differences in the roles and networks of genes in pre‐implantation embryos between species. Materials and methods To analyse the functions of SOX2 in lineage segregation and cell proliferation, loss‐ and gain‐of‐function studies were conducted in pig embryos using an overexpression vector and the CRISPR/Cas9 system. Then, we analysed the morphological features and examined the effect on the expression of downstream genes through immunocytochemistry and quantitative real‐time PCR. Results Our results showed that among the core pluripotent factors, only SOX2 was specifically expressed in the ICM. In SOX2‐disrupted blastocysts, the expression of the ICM‐related genes, but not OCT4, was suppressed, and the total cell number was also decreased. Likewise, according to real‐time PCR analysis, pluripotency‐related genes, excluding OCT4, and proliferation‐related genes were decreased in SOX2‐targeted blastocysts. In SOX2‐overexpressing embryos, the total blastocyst cell number was greatly increased but the ICM/TE ratio decreased. Conclusions Taken together, our results demonstrated that SOX2 is essential for ICM formation and cell proliferation in porcine early‐stage embryogenesis.

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“…The first insertion of transgenic material into mice embryos was achieved in 1974, and the passage of implanted genes to offspring in 1981. In 1987, the first method for genetically ablating genes was reported (reviewed: Saunders 2020 ), completing a suite of tools required for further probing the roles of iron-regulatory proteins suspected of playing important roles in the pathogenesis of Parkinson disease (Gerlach et al 1994 ).…”

Section: –2010: the Transgenic Toolbox Opensmentioning

confidence: 99%

A brief history of brain iron accumulation in Parkinson disease and related disorders

Double

2022

J Neural Transm

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Iron has a long and storied history in Parkinson disease and related disorders. This essential micronutrient is critical for normal brain function, but abnormal brain iron accumulation has been associated with extrapyramidal disease for a century. Precisely why, how, and when iron is implicated in neuronal death remains the subject of investigation. In this article, we review the history of iron in movement disorders, from the first observations in the early twentieth century to recent efforts that view extrapyramidal iron as a novel therapeutic target and diagnostic indicator.

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The History of Transgenesis

Cited by 13 publications

References 175 publications

Broad spectrum of CRISPR-induced edits in an embryonic lethal gene

Broad spectrum of CRISPR-induced edits in an embryonic lethal gene

SOX2 plays a crucial role in cell proliferation and lineage segregation during porcine pre‐implantation embryo development

A brief history of brain iron accumulation in Parkinson disease and related disorders

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