Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats

Aoshima, Takuya; Kobayashi, Yukari; Takagi, Hisayoshi; Iijima, Kenta; Takabayashi, Shuji

doi:10.1186/s12896-021-00723-5

Cited by 8 publications

(18 citation statements)

References 49 publications

(45 reference statements)

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“…However, the opposite results were recently achieved by Takabayashi et al [ 90 ] and Aoshima et al [ 141 ]. Takabayashi et al [ 90 ] performed i -GONAD using a solution containing 1 μM Scr7, RNP (1 μg/μL of Cas9 protein +30 µM of sgRNA), and 2 µg/µL of ssODN resulted in the production of 27 fetuses, of which only two (7%) had pigmented eyes.…”

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 70%

“…They named this technology “Technique for Animal Knockout system by Electroporation (TAKE)”. Since then, many GE rats have been produced using EP-based GE technology [ 54 , 90 , 91 , 95 , 111 , 117 , 121 , 125 , 131 , 140 , 141 , 144 ]. Notably, in vitro EP can also be applied to frozen mouse and rat zygotes for the efficient introduction of CRISPR components, without affecting the embryo viability or development [ 173 , 174 ].…”

Section: Delivery Methodsmentioning

confidence: 99%

“…They used a WT ssODN to exchange a single base pair in Tyr , thus reverting the albino allele back to the WT sequence and restoring a non-albino pigmentation pattern. A similar strategy using ssODN for the correction of mutated phenotypes in rats was also employed by others [ 64 , 90 , 91 , 106 , 114 , 125 , 126 , 127 , 129 , 141 , 142 ].…”

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

“…Notably, in the absence of Scr7, i -GONAD-mediated KI efficiency was as low as ~5%, suggesting that addition of Scr7 appeared not to affect i -GONAD-mediated KI efficiency in rats. Aoshima et al [ 141 ] examined whether commercially available KI-enhancing drugs (including Scr7, L755,507, RS-1, and HDR enhancer), all of which are known to enhance KI efficiency, can increase the KI efficiency using i -GONAD in rats. Aoshima et al [ 141 ] demonstrated that some drugs (e.g., Scr7, L755,507, and HDR enhancer) were found to be slightly effective in vivo, but their effects were not statistically significant when compared to those in the control group (without any KI-enhancing drugs).…”

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

“…Aoshima et al [ 141 ] constructed three gRNAs showing high efficiencies, ranging from 51% to 57%, based on the CHOPCHOP predictions [ 205 ]. These three gRNAs (termed crRNA1, crRNA2, and crRNA3) used here recognize the Tyr sequence containing a point mutation (G to A) and overlap with each other ( Figure 4 ).…”

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

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Recent Advances in the Production of Genome-Edited Rats

Nakamura

Inada

Takabayashi

2022

IJMS

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The rat is an important animal model for understanding gene function and developing human disease models. Knocking out a gene function in rats was difficult until recently, when a series of genome editing (GE) technologies, including zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the type II bacterial clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated Cas9 (CRISPR/Cas9) systems were successfully applied for gene modification (as exemplified by gene-specific knockout and knock-in) in the endogenous target genes of various organisms including rats. Owing to its simple application for gene modification and its ease of use, the CRISPR/Cas9 system is now commonly used worldwide. The most important aspect of this process is the selection of the method used to deliver GE components to rat embryos. In earlier stages, the microinjection (MI) of GE components into the cytoplasm and/or nuclei of a zygote was frequently employed. However, this method is associated with the use of an expensive manipulator system, the skills required to operate it, and the egg transfer (ET) of MI-treated embryos to recipient females for further development. In vitro electroporation (EP) of zygotes is next recognized as a simple and rapid method to introduce GE components to produce GE animals. Furthermore, in vitro transduction of rat embryos with adeno-associated viruses is potentially effective for obtaining GE rats. However, these two approaches also require ET. The use of gene-engineered embryonic stem cells or spermatogonial stem cells appears to be of interest to obtain GE rats; however, the procedure itself is difficult and laborious. Genome-editing via oviductal nucleic acids delivery (GONAD) (or improved GONAD (i-GONAD)) is a novel method allowing for the in situ production of GE zygotes existing within the oviductal lumen. This can be performed by the simple intraoviductal injection of GE components and subsequent in vivo EP toward the injected oviducts and does not require ET. In this review, we describe the development of various approaches for producing GE rats together with an assessment of their technical advantages and limitations, and present new GE-related technologies and current achievements using those rats in relation to human diseases.

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Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 70%

Section: Delivery Methodsmentioning

confidence: 99%

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

Section: Other Techniques and Factors To Modify The Rat Genomementioning

confidence: 99%

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Recent Advances in the Production of Genome-Edited Rats

Nakamura

Inada

Takabayashi

2022

IJMS

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Targeted insertion and reporter transgene activity at a gene safe harbor of the human blood fluke, Schistosoma mansoni

Ittiprasert,

Moescheid,

Chaparro

et al. 2023

Cell Reports Methods

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Superovulation with an anti-inhibin monoclonal antibody improves the reproductive performance of rat strains by increasing the pregnancy rate and the litter size

Mochida,

Morita,

Sasaoka

et al. 2024

Sci Rep

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Rats are multiparous rodents that have been used extensively in research; however, the low reproductive performance of some rat strains hampers the broader use of rats as a biomedical model. In this study, the possibility of increasing the litter size after natural mating in rats through superovulation using an anti-inhibin monoclonal antibody (AIMA) was examined. In outbred Wistar rats, AIMA increased the number of ovulated oocytes by 1.3-fold. AIMA did not affect fertilization and subsequent embryonic development, resulting in a 1.4-fold increase in litter size and a high pregnancy rate (86%). In contrast, conventional superovulation by eCG/hCG administration decreased the pregnancy rate to 6–40% and did not increase the litter size. In inbred Brown Norway rats, AIMA increased the litter size by 1.2-fold, and the pregnancy rate increased more than twice (86% versus 38% in controls). AIMA also increased the litter size by 1.5-fold in inbred Tokai High Avoiders and Fischer 344 rats. AIMA increased the efficiency of offspring production by 1.5-, 2.7-, 1.4-, and 1.4-fold, respectively, in the four rat strains. Thus, AIMA may consistently improve the reproductive performance through natural mating in rats, which could promote the use of AIMA in biomedical research.

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Modification of improved-genome editing via oviductal nucleic acids delivery (i-GONAD)-mediated knock-in in rats

Cited by 8 publications

References 49 publications

Recent Advances in the Production of Genome-Edited Rats

Recent Advances in the Production of Genome-Edited Rats

Targeted insertion and reporter transgene activity at a gene safe harbor of the human blood fluke, Schistosoma mansoni

Superovulation with an anti-inhibin monoclonal antibody improves the reproductive performance of rat strains by increasing the pregnancy rate and the litter size

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