Factors affecting production of transgenic rats by ICSI-mediated DNA transfer: Effects of sonication and freeze-thawing of spermatozoa, rat strains for sperm and oocyte donors, and different constructs of exogenous DNA

2005 ◽  
Vol 70 (4) ◽  
pp. 422-428 ◽  
Author(s):  
Masumi Hirabayashi ◽  
Megumi Kato ◽  
Ayako Ishikawa ◽  
Ryosuke Kaneko ◽  
Takeshi Yagi ◽  
...  
Keyword(s):  
Dna Transfer ◽  
Transfer Effects ◽  
Transgenic Rats ◽  
Rat Strains ◽  
Exogenous Dna ◽  
Factors Affecting ◽  
Oocyte Donors

scholarly journals 317 FACTORS AFFECTING PRODUCTION EFFICIENCY OF TRANSGENIC RATS BY ICSI-MEDIATED DNA TRANSFER

2005 ◽  
Vol 17 (2) ◽  
pp. 309
Author(s):  
M. Kato ◽  
S. Hochi ◽  
M. Hirabayashi
Keyword(s):  
Production Efficiency ◽  
Dna Transfer ◽  
Egfp Expression ◽  
Transgenic Rat ◽  
Sprague Dawley ◽  
Transgenic Rats ◽  
Rat Strains ◽  
Exogenous Dna ◽  
Factors Affecting ◽  
Rat Offspring

Successful DNA transfer via intracytoplasmic sperm injection (ICSI) was first reported in mice (Perry et al. 1999, Science 284, 1180), and was recently extended to rats (Kato et al. 2004, Mol. Reprod. Dev. 69, 153). In the present study, factors affecting the production efficiency of transgenic rats by the ICSI-mediated DNA transfer were investigated. Cauda epididymal spermatozoa from Sprague-Dawley rats were sonicated (SO) and/or frozen-thawed (FT) for tail-cutting and membrane-disrupting. The sperm heads were exposed for 1 min to different concentrations (0.02–2.5 ng/μL) of 3.0 kb EGFP DNA solution, and then microinjected into denuded F1 (Donryu × LEW) rat oocytes. The optimal concentration of EGFP DNA was 0.1 ng/μL, as determined by the in vitro developmental competence into morulae/blastocysts and the EGFP expression of the ICSI oocytes. The presumptive 1- or 2-cell stage zygotes were transferred into oviducts of pseudopregnant Wistar females, and the presence of EGFP DNA in the offspring was examined by fluorescence under the 480 nm UV light. The production efficiency of transgenic rat offspring was 2.8% (2/71 zygotes transferred), 1.6% (1/63), and 3.3% (2/61) in the oocytes into which SO-, FT-, and SO+ FT-treated sperm heads were injected, respectively. The founder transgenic rats carrying EGFP DNA transmitted the transgenes to their progeny according to the Mendelian fashion (43.8–54.8%), suggesting the stable incorporation of the transgenes into rat genomes. Four rat strains (F344, LEW, Donryu, and Sprague-Dawley) were compared for their suitability as sperm/oocyte donors in the production of transgenic rats by ICSI with SO + FT-treated and 0.1 ng/μL EGFP DNA-exposed sperm heads. The production efficiency of the transgenic rats in the Sprague-Dawley strain (8.2%, 8/98) was significantly higher than that in LEW strain (0.9%, 1/114), while those in F344 (4.3%, 4/92) and Donryu (4.4%, 5/114) strains were intermediate. Attempts were made to introduce three other DNA constructs (5.0 kb plasmid and 208 kb BAC, both with Fyn gene, and 186 kb BAC with Svet1/IRES-Cre gene) into rat genomes by ICSI with SO + FT-treated and 0.1 ng/μL DNA-exposed sperm heads. PCR analysis showed that the Fyn, Fyn/BAC, and Svet1/IRES-Cre DNA constructs were successfully introduced into Sprague-Dawley rat offspring via ICSI, with production efficiencies of 2.8% (3/109), 0.9% (1/109), and 2.4% (3/125), respectively. These results indicate that transgenic rats can be produced by ICSI-mediated DNA transfer using the various types of exogenous DNA and rat strains with different genetic backgrounds.

Effectiveness of exogenous DNA transfer to chicken embryo cells in vitro and in vivo using retroviral vectors

2007 ◽  
Vol 33 (3) ◽  
pp. 180-182 ◽  
Author(s):  
N. A. Volkova ◽  
A. O. Tulyakova ◽  
L. A. Volkova ◽  
N. A. Zinov’eva ◽  
L. K. Ernst ◽  
...  
Keyword(s):  
Chicken Embryo ◽  
Retroviral Vectors ◽  
Dna Transfer ◽  
Exogenous Dna ◽  
Embryo Cells

Efficient and simple production of transgenic mice and rabbits using the new DMSO-sperm mediated exogenous DNA transfer method

2006 ◽  
Vol 73 (5) ◽  
pp. 589-594 ◽  
Author(s):  
Wei Shen ◽  
Lan Li ◽  
Qingjie Pan ◽  
Lingjiang Min ◽  
Huansheng Dong ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Dna Transfer ◽  
Exogenous Dna ◽  
Transfer Method ◽  
Simple Production

scholarly journals Factors Affecting the Producing Efficiency of Transgenic Rats

1997 ◽  
Vol 43 (5) ◽  
pp. j15-j18 ◽  
Author(s):  
Masumi HIRABAYASHI ◽  
Ri-ichi TAKAHASHI ◽  
Kazumi ITO ◽  
Kunihiko KODAIRA ◽  
Takashige SUZUKI ◽  
...  
Keyword(s):  
Transgenic Rats ◽  
Factors Affecting

Production of transgenic rats by ooplasmic injection of spermatogenic cells exposed to exogenous DNA: A preliminary study

2004 ◽  
Vol 69 (2) ◽  
pp. 153-158 ◽  
Author(s):  
Megumi Kato ◽  
Ayako Ishikawa ◽  
Ryosuke Kaneko ◽  
Takeshi Yagi ◽  
Shinichi Hochi ◽  
...  
Keyword(s):  
Spermatogenic Cells ◽  
Transgenic Rats ◽  
Exogenous Dna ◽  
Preliminary Study

scholarly journals Desiccation does not drastically increase the accessibility of exogenous DNA to nuclear genomes: Evidence from the frequency of endosymbiotic DNA transfer

2020 ◽  
Author(s):  
Xi-Xi Li ◽  
Cheng Fang ◽  
Jun-Peng Zhao ◽  
Xiao-Yu Zhou ◽  
Zhihua Ni ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Nuclear Genome ◽  
Dna Transfer ◽  
Dna Double Strand Breaks ◽  
End Joining ◽  
Exogenous Dna ◽  
Bdelloid Rotifers ◽  
Significant Difference ◽  
Non Homologous End Joining ◽  
High Level

Abstract Background: Although horizontal gene transfer (HGT) is a widely accepted force in the evolution of prokaryotic genomes, its role in the evolution of eukaryotic genomes remains hotly debated. Some bdelloid rotifers that are resistant to extreme desiccation and radiation undergo a very high level of HGT, whereas a similar report of another desiccation-resistant invertebrate, the tardigrade, remains controversial. Overall, the DNA double-strand breaks (DSBs) induced by prolonged desiccation have been postulated to open a gateway to the nuclear genome for exogenous DNA integration and thus to facilitate the HGT process, thereby enhancing the rate of endosymbiotic DNA transfer (EDT). Results: We first surveyed the abundance of nuclear mitochondrial DNAs (NUMTs) and nuclear plastid DNAs (NUPTs) in five eukaryotes that are highly resistant to desiccation: the bdelloid rotifers Adineta vaga and Adineta ricciae , the tardigrade Ramazzottius varieornatus , and the resurrection plants Dorcoceras hygrometricum and Selaginella tamariscina . Excessive NUMTs or NUPTs were not detected. Furthermore, we compared 24 groups of desiccation-tolerant organisms with their relatively less desiccation-tolerant relatives but did not find a significant difference in NUMT/NUPT contents. Conclusions: Desiccation may induce DSBs, but it is unlikely to dramatically increase the frequency of exogenous sequence integration in most eukaryotes. The capture of exogenous DNA sequences is possible only when DSBs are repaired through a subtype of non-homologous end joining, named alternative end joining (alt-EJ). Due to the deleterious effects of the resulting insertion mutations, alt-EJ is less frequently initiated than other mechanisms.

Methodological approaches for vitrification of bovine oocytes

Zygote ◽  
2020 ◽  
pp. 1-11
Author(s):  
Linda Dujíčková ◽  
Alexander V. Makarevich ◽  
Lucia Olexiková ◽  
Elena Kubovičová ◽  
František Strejček
Keyword(s):  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Oocyte Vitrification ◽  
Factors Affecting ◽  
Oocyte Stage ◽  
Follicle Size ◽  
Oocyte Donors ◽  
Methodological Approaches ◽  
Bovine Oocytes

Summary Numerous factors affect vitrification success and post-thaw development of oocytes after in vitro fertilization. Therefore, elaboration of an optimal methodology ensuring higher cryotolerance of oocytes and subsequent blastocyst yield is still of great interest. This paper describes and evaluates critical factors affecting the success of oocyte vitrification. In particular, an appropriate oocyte stage such as maturation status (germinal vesicle stage, metaphase II stage), presence/absence of cumulus cells before vitrification, and the effect of follicle size, as well as different culture systems and media for in vitro production of embryos, the types and concentrations of cryoprotectants, and cooling and warming rates at vitrification are considered. Special attention is paid to various cryocarriers used for low-volume vitrification, which ensures safe storage of oocytes/embryos in liquid nitrogen and their successful post-thaw recovery. At the end, we focussed on how age of oocyte donors (heifers, cows) influences post-thaw development. This review summarizes results of recently published studies describing different methodologies of cryopreservation and post-thaw oocyte development with the main focus on vitrification of bovine oocytes.

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