scholarly journals Possible Mechanisms for the Testis-Mediated Gene Transfer as a New Method for Producing Transgenic Animals.

1999 ◽  
Vol 45 (1) ◽  
pp. 37-42 ◽  
Author(s):  
Kyu-Tae Chang ◽  
Akihiro Ikeda ◽  
Katsuhiko Hayashi ◽  
Yasufumi Furuhata ◽  
Makoto Bannai ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Transgenic Animals ◽  
New Method

Sonication: A new method for gene transfer to plants

1992 ◽  
Vol 85 (2) ◽  
pp. 230-234 ◽  
Author(s):  
Morten Joersbo ◽  
Janne Brunstedt
Keyword(s):  
Gene Transfer ◽  
New Method

Chromosomal gene transfer: prospects of a new method for mapping closely linked human genes

1976 ◽  
Vol 16 (1-5) ◽  
pp. 405-408 ◽  
Author(s):  
K. Willecke ◽  
P.J. Davies ◽  
T. Reber
Keyword(s):  
Gene Transfer ◽  
New Method ◽  
Chromosomal Gene ◽  
Human Genes

scholarly journals Production of transgenic piglets using ICSI–sperm-mediated gene transfer in combination with recombinase RecA

Reproduction ◽  
2010 ◽  
Vol 140 (2) ◽  
pp. 259-272 ◽  
Author(s):  
Francisco A García-Vázquez ◽  
Salvador Ruiz ◽  
Carmen Matás ◽  
M José Izquierdo-Rico ◽  
Luis A Grullón ◽  
...  
Keyword(s):  
Gene Transfer ◽  
High Efficiency ◽  
Fluorescent Protein ◽  
Reactive Oxygen Species Generation ◽  
Reca Protein ◽  
Transgenic Animals ◽  
Membrane Lipid ◽  
Transgenic Pigs ◽  
Exogenous Dna

Sperm-mediated gene transfer (SMGT) is a method for the production of transgenic animals based on the intrinsic ability of sperm cells to bind and internalize exogenous DNA molecules and to transfer them into the oocyte at fertilization. Recombinase-A (RecA) protein-coated exogenous DNA has been used previously in pronuclear injection systems increasing integration into goat and pig genomes. However, there are no data regarding transgene expression after ICSI. Here, we set out to investigate whether the expression of transgenic DNA in porcine embryos is improved by recombinase-mediated DNA transfer and if it is possible to generate transgenic animals using this methodology. Different factors which could affect the performance of this transgenic methodology were analyzed by studying 1) the effect of the presence of exogenous DNA and RecA protein on boar sperm functionality; 2) the effect of recombinase RecA on in vitro enhanced green fluorescent protein (EGFP)-expressing embryos produced by ICSI or IVF; and 3) the efficiency of generation of transgenic piglets by RecA-mediated ICSI. Our results suggested that 1) the presence of exogenous DNA and RecA–DNA complexes at 5 μg/ml did not affect sperm functionality in terms of motility, viability, membrane lipid disorder, or reactive oxygen species generation; 2) EGFP-expressing embryos were obtained with a high efficiency using the SMGT–ICSI technique in combination with recombinase; however, the use of IVF system did not result in any fluorescent embryos; and 3) transgenic piglets were produced by this methodology. To our knowledge, this is the first time that transgenic pigs have been produced by ICSI-SGMT and a recombinase.

Sonication: A new method for gene transfer to plants

1992 ◽  
Vol 85 (2) ◽  
pp. 230-234 ◽  
Author(s):  
Morten Joersbo ◽  
Janne Brunstedt
Keyword(s):  
Gene Transfer ◽  
New Method

scholarly journals Fluorescence-Based Detection of Natural Transformation in Drug-ResistantAcinetobacter baumannii

2018 ◽  
Vol 200 (19) ◽  
Author(s):  
Anne-Sophie Godeux ◽  
Agnese Lupo ◽  
Marisa Haenni ◽  
Simon Guette-Marquet ◽  
Gottfried Wilharm ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Natural Transformation ◽  
Multidrug Resistant ◽  
New Method ◽  
Content Type ◽  
Transfer Mechanisms ◽  
Multiple Antibiotics

ABSTRACTAcinetobacter baumanniiis a nosocomial agent with a high propensity for developing resistance to antibiotics. This ability relies on horizontal gene transfer mechanisms occurring in theAcinetobactergenus, including natural transformation. To study natural transformation in bacteria, the most prevalent method uses selection for the acquisition of an antibiotic resistance marker in a target chromosomal locus by the recipient cell. Most clinical isolates ofA. baumanniiare resistant to multiple antibiotics, limiting the use of such selection-based methods. Here, we report the development of a phenotypic and selection-free method based on flow cytometry to detect transformation events in multidrug-resistant (MDR) clinicalA. baumanniiisolates. To this end, we engineered a translational fusion between the abundant and conservedA. baumanniinucleoprotein (HU) and the superfolder green fluorescent protein (sfGFP). The new method was benchmarked against the conventional antibiotic selection-based method. Using this new method, we investigated several parameters affecting transformation efficiencies and identified conditions of transformability one hundred times higher than those previously reported. Using optimized transformation conditions, we probed natural transformation in a set of MDR clinical and nonclinical animalA. baumanniiisolates. Regardless of their origin, the majority of the isolates displayed natural transformability, indicative of a conserved trait in the species. Overall, this new method and optimized protocol will greatly facilitate the study of natural transformation in the opportunistic pathogenA. baumannii.IMPORTANCEAntibiotic resistance is a pressing global health concern with the rise of multiple and panresistant pathogens. The rapid and unfailing resistance to multiple antibiotics of the nosocomial agentAcinetobacter baumannii, notably to carbapenems, prompt to understand the mechanisms behind acquisition of new antibiotic resistance genes. Natural transformation, one of the horizontal gene transfer mechanisms in bacteria, was only recently described inA. baumanniiand could explain its ability to acquire resistance genes. We developed a reliable method to probe and study natural transformation mechanism inA. baumannii. More broadly, this new method based on flow cytometry will allow experimental detection and quantification of horizontal gene transfer events in multidrug-resistantA. baumannii.

scholarly journals Retrotransposon-mediated Gene Transfer for Animal Cells

2021 ◽  
Vol 333 ◽  
pp. 07002
Author(s):  
Feiyang Zheng ◽  
Yoshinori Kawabe ◽  
Mai Murakami ◽  
Mamika Takahashi ◽  
Shoichiro Yoshida ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Culture Medium ◽  
Transgenic Animals ◽  
Open Reading Frames ◽  
Pcr Analysis ◽  
Animal Cells ◽  
Cmv Promoter ◽  
Pharmaceutical Protein ◽  
Cell Clones ◽  
Copy And Paste

Gene delivery methods for animal cells are one of the most important tools in biotechnology fields such as pharmaceutical protein production, generation of transgenic animals and gene therapy. Because retrotransposons can move their own sequences to new genomic locations by a “copy-and-paste” process known as retrotransposition, we attempted to develop a novel gene transfer system based on retrotransposon. A full-length long interspersed element-1 (LINE-1) contains a 5’ untranslated region (5’UTR), two non-overlapping open reading frames (ORFs) separated by a short inter-ORF sequence, and a 3’UTR terminating in an adenosine-rich tract. We constructed a LINE-1 vector plasmid including components necessary for retrotransposition. An intron-disrupted Neo reporter gene and a scFv-Fc expression unit under the control of CMV promoter were added into 3’UTR in order to evaluate retrotransposition and express scFv-Fc. CHO-K1 cells transfected with the plasmids were screened with G418. The established cell clones produced scFv-Fc proteins in the culture medium. To control retrotransposition steadily, we also established retrotransposon systems that supply ORF2 or ORF1–2 separately. Genomic PCR analysis revealed that transgene sequences derived from the LINE-1 vector were positive in all clones. All the clones tested produced scFv-Fc in the culture medium.

Sign in / Sign up

Export Citation Format

Share Document