Advances in Genome Editing and Application to the Generation of Genetically Modified Rat Models

Chenouard, Vanessa; Remy, Séverine; Tesson, Laurent; Ménoret, Séverine; Ouisse, Laure-Hélène; Chérifi, Yacine; Anegón, Ignacio

doi:10.3389/fgene.2021.615491

Cited by 24 publications

(13 citation statements)

References 387 publications

(398 reference statements)

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“…ITGA5, MMP12). Strategies exist in the rat for testing in vivo roles of candidates using global and conditional genome editing ( 39 ) and trophoblast cell-specific lentiviral mediated gene manipulation ( 40, 41 ). Dysregulation of candidate genes may directly affect the development and function of the invasive trophoblast cell lineage and indirectly affect uterine cell dynamics at the uterine-placental interface.…”

Section: Discussionmentioning

confidence: 99%

Conservation at the uterine-placental interface

Scott

Jain

et al. 2022

Preprint

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The hemochorial placentation site is characterized by a dynamic interplay between trophoblast cells and maternal cells. These cells cooperate to establish an interface required for nutrient delivery to promote fetal growth. In the human, trophoblast cells penetrate deep into the uterus. This is not a consistent feature of hemochorial placentation and has hindered the establishment of suitable animal models. The rat represents an intriguing model for investigating hemochorial placentation with deep trophoblast cell invasion. In this study, we used single cell RNA sequencing to characterize the transcriptome of the invasive trophoblast cell lineage, as well as other cell populations within the rat uterine-placental interface during early (gestation day, gd, 15.5) and late (gd 19.5) stages of intrauterine trophoblast cell invasion. We identified a robust set of transcripts that define invasive trophoblast cells, as well as transcripts that distinguished endothelial, smooth muscle, natural killer, and macrophage cells. Invasive trophoblast, immune, and endothelial cell populations exhibited distinct spatial relationships within the uterine-placental interface. Furthermore, the maturation stage of invasive trophoblast cell development could be determined by assessing gestation-stage dependent changes in transcript expression. Finally, and most importantly, expression of a prominent subset of rat invasive trophoblast cell transcripts is conserved in the invasive extravillous trophoblast cell lineage of the human placenta. These findings provide foundational data to identify and interrogate key conserved regulatory mechanisms essential for development and function of an important compartment within the hemochorial placentation site that is essential for a healthy pregnancy.

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Section: Discussionmentioning

confidence: 99%

Conservation at the uterine-placental interface

Scott

Jain

et al. 2022

Preprint

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“…Nonetheless, it is still difficult to use GONAD to introduce larger mutations requiring a long DNA donor (>2 kb). If even larger knockins need to be engineered, other methods such as microinjection and CRISPR-READI may be more suitable (Chen et al, 2019;Chenouard et al, 2021).…”

Section: Backg Round and Fe Ature Smentioning

confidence: 99%

GONAD: A new method for germline genome editing in mice and rats

et al. 2021

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Recent advances in the CRISPR/Cas9 system have demonstrated it to be an efficient gene-editing technology for various organisms. Laboratory mice and rats are widely used as common models of human diseases; however, the current standard method to create genome-engineered animals is laborious and involves three major steps: isolation of zygotes from females, ex vivo micromanipulation of zygotes, and implantation into pseudopregnant females. To circumvent this, we recently developed a novel method named Genome-editing via Oviductal Nucleic Acids Delivery (GONAD). This method does not require the ex vivo handling of embryos; instead, it can execute gene editing with just one step, via the delivery of a genome-editing mixture into embryos in the oviduct, by electroporation. Here, we present a further improvement of GONAD that is easily applicable to both mice and rats. It is a rapid, low-cost, and ethical approach fulfilling the 3R principles of animal experimentation: Reduction, Replacement, and Refinement. This method has been reconstructed and renamed as "improved GONAD (i-GONAD)" for mice, and "rat improved GONAD (rGONAD)" for rats.

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“… 28 The complement levels in rats and humans are comparable, whereas in most inbred mice they are very low. 29 , 30 In this context, we used rats to construct a Trdmt1 gene knockout model and adopted LPS‐induced inflammation to study the function of Trdmt1 in inflammation progression. Our data showed that Trdmt1 deletion made rats more vulnerable to LPS treatment.…”

Section: Introductionmentioning

confidence: 99%

“…For example, rats share more immune characteristics with humans than mice do 28 . The complement levels in rats and humans are comparable, whereas in most inbred mice they are very low 29,30 . In this context, we used rats to construct a Trdmt1 gene knockout model and adopted LPS‐induced inflammation to study the function of Trdmt1 in inflammation progression.…”

Section: Introductionmentioning

confidence: 99%

TRDMT1 exhibited protective effects against LPS‐induced inflammation in rats through TLR4‐NF‐κB/MAPK‐TNF‐α pathway

Zhang

et al. 2022

Anim Models and Exp Med

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Background Inflammation is a complex physiological and pathological process. Although many types of inflammation are well characterized, their physiological functions are largely unknown. tRNA aspartic acid methyltransferase 1 (TRDMT1) has been implicated as a stress‐related protein, but its intrinsic biological role is unclear. Methods We constructed a Trdmt1 knockout rat and adopted the LPS‐induced sepsis model. Survival curve, histopathological examination, expression of inflammatory factors, and protein level of TLR4 pathway were analyzed. Results Trdmt1 deletion had no obvious impact on development and growth. Trdmt1 deletion slightly increased the mortality during aging. Our data showed that Trdmt1 strongly responded in LPS‐treated rats, and Trdmt1 knockout rats were vulnerable to LPS treatment with declined survival rate. We also observed more aggravated tissue damage and more cumulative functional cell degeneration in LPS‐treated knockout rats compared with control rats. Further studies showed upregulated TNF‐α level in liver, spleen, lung, and serum tissues, which may be explained by enhanced p65 and p38 phosphorylation. Conclusions Our data demonstrated that Trdmt1 plays a protective role in inflammation by regulating the TLR4‐NF‐κB/MAPK‐TNF‐α pathway. This work provides useful information to understand the TRDMT1 function in inflammation.

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Advances in Genome Editing and Application to the Generation of Genetically Modified Rat Models

Cited by 24 publications

References 387 publications

Conservation at the uterine-placental interface

Conservation at the uterine-placental interface

GONAD: A new method for germline genome editing in mice and rats

TRDMT1 exhibited protective effects against LPS‐induced inflammation in rats through TLR4‐NF‐κB/MAPK‐TNF‐α pathway

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