scholarly journals SAT-111 Rat genome editing by rGONAD (rat Genome-editing via Oviductal Nucleic Acids Delivery) method

2019 ◽  
Vol 4 (7) ◽  
pp. S52
Author(s):  
M. Matsuyama ◽  
T. Koyano ◽  
T. Kobayashi ◽  
M. Namba ◽  
M. Fukushima
2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Shuji Takabayashi ◽  
Takuya Aoshima ◽  
Katsuya Kabashima ◽  
Kazushi Aoto ◽  
Masato Ohtsuka ◽  
...  

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Takuya Aoshima ◽  
Yukari Kobayashi ◽  
Hisayoshi Takagi ◽  
Kenta Iijima ◽  
Masahiro Sato ◽  
...  

Abstract Background Improved genome-editing via oviductal nucleic acids delivery (i-GONAD) is a new technology that facilitates in situ genome-editing of mammalian zygotes exiting the oviductal lumen. The i-GONAD technology has been developed for use in mice, rats, and hamsters; however, oligonucleotide (ODN)-based knock-in (KI) is more inefficient in rats than mice. To improve the efficiency of i-GONAD in rats we examined KI efficiency using three guide RNAs (gRNA), crRNA1, crRNA2 and crRNA3. These gRNAs recognize different portions of the target locus, but also overlap each other in the target locus. We also examined the effects of commercially available KI -enhancing drugs (including SCR7, L755,507, RS-1, and HDR enhancer) on i-GONAD-mediated KI efficiency. Results The KI efficiency in rat fetuses generated after i-GONAD with crRNA2 and single-stranded ODN was significantly higher (24%) than crRNA1 (5%; p < 0.05) or crRNA3 (0%; p < 0.01). The KI efficiency of i-GONAD with triple gRNAs was 11%. These findings suggest that KI efficiency largely depends on the type of gRNA used. Furthermore, the KI efficiency drugs, SCR7, L755,507 and HDR enhancer, all of which are known to enhance KI efficiency, increased KI efficiency using the i-GONAD with crRNA1 protocol. In contrast, only L755,507 (15 μM) increased KI efficiency using the i-GONAD with crRNA2 protocol. None of them were significantly different. Conclusions We attempted to improve the KI efficiency of i-GONAD in rats. We demonstrated that the choice of gRNA is important for determining KI efficiency and insertion and deletion rates. Some drugs (e.g. SCR7, L755,507 and HDR enhancer) that are known to increase KI efficiency in culture cells were found to be effective in i-GONAD in rats, but their effects were limited.


2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Gou Takahashi ◽  
Channabasavaiah B Gurumurthy ◽  
Kenta Wada ◽  
Hiromi Miura ◽  
Masahiro Sato ◽  
...  

2016 ◽  
Vol 11 (1) ◽  
Author(s):  
Masahiro Sato ◽  
Masato Ohtsuka ◽  
Satoshi Watanabe ◽  
Channabasavaiah B. Gurumurthy

1970 ◽  
Vol 8 (2) ◽  
pp. 50-57
Author(s):  
Jemal Dilebo

Mesoporous silica nanoparticles (MSN) have been explored for the delivery of small molecule drugs, antigens, and nucleic acids because of their large surface area, pore volume, amenability of their surface for functionalization, stable mesoporous structure, and biocompatibility.  Biomoecules loading capacitites,  release and target cell accumulation efficiencies have been improved for both antigen and nucleic acid delivery by the synthesis of large-pore MSN, dendritic MSN, hollow-core MSN, and multifunctional MSN. This article overview the major advances in the use of MSN for delivery of antigens and therapeutic nucleic acids such as DNA, siRNA, and miRNA aimed for treatment of various diseases.       


2015 ◽  
pp. 285-336
Author(s):  
Erea Borrajo ◽  
Anxo Vidal ◽  
Maria J. Alonso ◽  
Marcos Garcia-Fuentes

2020 ◽  
Vol 3 (5) ◽  
pp. 2779-2795 ◽  
Author(s):  
Jin Huang ◽  
Wenjie Ma ◽  
Huanhuan Sun ◽  
Huizhen Wang ◽  
Xiaoxiao He ◽  
...  

MedChemComm ◽  
2010 ◽  
Vol 1 (1) ◽  
pp. 76 ◽  
Author(s):  
Guilhem Godeau ◽  
Hélène Arnion ◽  
Christophe Brun ◽  
Cathy Staedel ◽  
Philippe Barthélémy

2018 ◽  
Vol 6 (3) ◽  
pp. 438-451 ◽  
Author(s):  
Yuwei Zhu ◽  
Zhiwei Huang

Abstract Clustered regularly interspaced short palindromic repeats (CRISPR) and accompanying CRISPR-associated (Cas) proteins provide RNA-guided adaptive immunity for prokaryotes to defend themselves against viruses. The CRISPR-Cas systems have attracted much attention in recent years for their power in aiding the development of genome editing tools. Based on the composition of the CRISPR RNA-effector complex, the CRISPR-Cas systems can be divided into two classes and six types. In this review, we summarize recent advances in the structural biology of the CRISPR-Cas-mediated genome editing tools, which helps us to understand the mechanism of how the guide RNAs assemble with diverse Cas proteins to cleave target nucleic acids.


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