scholarly journals Production of Transgenic Goats by Sperm-mediated Exogenous DNA Transfer Method

2009 ◽  
Vol 23 (1) ◽  
pp. 33-40 ◽  
Author(s):  
Yongju Zhao ◽  
Hong Wei ◽  
Yong Wang ◽  
Lingbin Wang ◽  
Mingju Yu ◽  
...  
Keyword(s):  
Dna Transfer ◽  
Exogenous Dna ◽  
Transgenic Goats ◽  
Transfer Method

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

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

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.

Contact Transfer Method: DNA Transfer from Polyacrylamide Gel

1995 ◽  
Vol 224 (2) ◽  
pp. 611-612 ◽  
Author(s):  
M. Takeuchi ◽  
H. Fujisawa
Keyword(s):  
Polyacrylamide Gel ◽  
Dna Transfer ◽  
Transfer Method

scholarly journals Highly Efficient CRISPR/Cas9 System in Plasmodium Falciparum Using Cas9-expressing Parasites and a Linear Donor Template

2021 ◽  
Author(s):  
Tsubasa Nishi ◽  
Naoaki Shinzawa ◽  
Masao Yuda ◽  
Shiroh Iwanaga
Keyword(s):  
Plasmodium Falciparum ◽  
High Efficiency ◽  
Standard Technique ◽  
Dna Transfer ◽  
Drug Selection ◽  
Technical Problems ◽  
Highly Efficient ◽  
Transgenic Parasites ◽  
Transfer Method ◽  
Donor Template

Abstract The current CRISPR/Cas9 system for Plasmodium falciparum suffers from technical problems caused by plasmid constructs, such as delays in establishing transgenic parasites during drug selection and unexpected integration of circular donor DNA by single-crossover recombination. Although these problems can be solved by using linear donor templates, such an approach requires highly efficient introduction of DNA and rapid completion of recombination because linear DNA is easily lost from the parasites during multiplication. Here, we overcame these problems by developing a highly efficient DNA transfer method and Cas9-expressing parasites. Using our new CRISPR/Cas9 system, transgenic parasites were established in two weeks without any unexpected recombination or off-target mutations. Furthermore, with our system, two genes on different chromosomes were successfully modified in one transfection. Because of its high efficiency and robustness, our new CRISPR/Cas9 system will become a standard technique for genetic engineering of P. falciparum and dramatically advance studies of this parasite.

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 in another desiccation-resistant invertebrate, the tardigrade, the pattern does not exist. 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.

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.

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