Lentiviral Transgenesis

2009 ◽  
pp. 391-405 ◽  
Author(s):  
Alexander Pfeifer ◽  
Andreas Hofmann
Keyword(s):  
Lentiviral Transgenesis

Characterisation of eGFP-transgenic BALB/c mouse strain established by lentiviral transgenesis

2009 ◽  
Vol 19 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Krisztián Kvell ◽  
Tamás Czömpöly ◽  
László Hiripi ◽  
Péter Balogh ◽  
József Kóbor ◽  
...  
Keyword(s):  
Mouse Strain ◽  
Lentiviral Transgenesis

Response to Whitelaw: Lentiviral transgenesis in livestock

2004 ◽  
Vol 22 (4) ◽  
pp. 159-160 ◽  
Author(s):  
Alexander Pfeifer ◽  
Andreas Hofmann ◽  
Barbara Kessler ◽  
Eckhard Wolf
Keyword(s):  
Lentiviral Transgenesis

New technology for an old favorite: lentiviral transgenesis and RNAi in rats

2007 ◽  
Vol 16 (5) ◽  
pp. 571-580 ◽  
Author(s):  
Christina Tenenhaus Dann
Keyword(s):  
New Technology ◽  
Lentiviral Transgenesis

Lentiviral transgenesis in mice via a simple method of viral concentration

2016 ◽  
Vol 86 (6) ◽  
pp. 1427-1435 ◽  
Author(s):  
Pei-Hsun Cheng ◽  
Yu-Fan Chang ◽  
Su-Han Mao ◽  
Hsiu-Lien Lin ◽  
Chuan-Mu Chen ◽  
...  
Keyword(s):  
Simple Method ◽  
Lentiviral Transgenesis

scholarly journals Restriction enzyme mediated integration and FIV lentiviral transgenesis applied to amphibians

2014 ◽  
Vol 8 (1) ◽  
pp. 1-11
Author(s):  
Hull Kaitlyn ◽  
Hodgson Dee ◽  
N. Petitte James ◽  
Clark Bob ◽  
Hickok, Tim ◽  
...  
Keyword(s):  
Restriction Enzyme ◽  
Lentiviral Transgenesis

Lentiviral transgenesis in livestock

2010 ◽  
Vol 20 (3) ◽  
pp. 441-442 ◽  
Author(s):  
Simon Lillico ◽  
Douglas Vasey ◽  
Tim King ◽  
Bruce Whitelaw
Keyword(s):  
Lentiviral Transgenesis

Lentiviral vectors: are they the future of animal transgenesis?

2007 ◽  
Vol 31 (2) ◽  
pp. 159-173 ◽  
Author(s):  
Frank Park
Keyword(s):  
Transgene Expression ◽  
High Efficiency ◽  
Lentiviral Vector ◽  
Transgenic Animals ◽  
Lentiviral Vectors ◽  
Vector System ◽  
Gene Therapy Vector ◽  
Advantages And Disadvantages ◽  
Lentiviral Transgenesis ◽  
Alternative Approaches

Lentiviral vectors have become a promising new tool for the establishment of transgenic animals and the manipulation of the mammalian genome. While conventional microinjection-based methods for transgenesis have been successful in generating small and large transgenic animals, their relatively low transgenic efficiency has opened the door for alternative approaches, including lentiviral vectors. Lentiviral vectors are an appealing tool for transgenesis in part because of their ability to incorporate into genomic DNA with high efficiency, especially in cells that are not actively dividing. Lentiviral vector-mediated transgene expression can also be maintained for long periods of time. Recent studies have documented high efficiencies for lentiviral transgenesis, even in animal species and strains, such as NOD/ scid and C57Bl/6 mouse, that are very difficult to manipulate using the standard transgenic techniques. These advantages of the lentiviral vector system have broadened its use as a gene therapy vector to additional applications that include transgenesis and knockdown functional genetics. This review will address the components of the lentiviral vector system and recent successes in lentiviral transgenesis using both male- and female-derived pluripotent cells. The advantages and disadvantages of lentiviral transgenesis vs. other approaches to produce transgenic animals will be compared with regard to efficiency, the ability to promote persistent transgene expression, and the time necessary to generate a sufficient number of animals for phenotyping.

372 LASER-ASSISTED LENTIVIRAL TRANSGENESIS IN BOVINE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 293
Author(s):  
S. Ewerling ◽  
A. Hofmann ◽  
R. Klose ◽  
K. Rink ◽  
G. Brem ◽  
...  
Keyword(s):  
Incubation Time ◽  
Cell Mass ◽  
Pulse Length ◽  
Egfp Expression ◽  
Virus Concentration ◽  
Virus Suspension ◽  
Cleavage Rate ◽  
Lentiviral Transgenesis ◽  
Maximum Humidity

Lentiviral vectors have been shown to be a powerful tool to create transgenic livestock (Hofmann et al. 2003 EMBO Rep. 4, 1054-1060; Hofmann et al. 2004 Biol. Reprod. 71, 405-409). Due to their inability to pass the zona pellucida (ZP) by themselves, viral particles were delivered by subzonal microinjection of oocytes or zygotes. Here we show that an artificial opening of the ZP, generated by an infrared 1.48 �m microsurgical diode laser (OCTAX Laser Shot", kindly provided by MTG, Altdorf, Germany) and subsequent culture of oocytes in virus suspension, was sufficient to allow virus integration and expression of eGFP. The ZP of denuded in vitro-matured bovine oocytes was microdrilled with 2 to 4 laser shots of 3.0-ms pulse length each to create an opening of at least 40 �m. Oocytes were transferred to microdroplets (20 �L of Fert Talp) containing the virus suspension (lentiviral vector containing the eGFP reporter gene under the control of the human phosphoglycerate-kinase 1 promoter). After incubation for 2 or 4 h (5% CO2, maximum humidity, 39�C), oocytes were washed thoroughly in fresh droplets of medium to remove excessive virus. Sperm was added (0.5 � 106/mL) for 8 h. In vitro culture of embryos (5% CO2, 5% O2, maximum humidity, 39�C) was continued in culture medium (SOF) under oil for up to 8 days. Cleavage rate and number of blastocysts were determined on Days 2 and 8, respectively. Expression of eGFP was analyzed on Day 8 by fluorescence microscopy using an eGFP-specific filter with UV light source under an inverted microscope. In total, 267 oocytes were microdrilled. The range of optimal virus concentration was analyzed by titration. Seven different concentrations in a range of 1.25 � 106 to 5 � 107 IU/mL were used at an incubation time of 4 h. In one experiment using a high virus concentration, the incubation time was reduced to 2 h. Control groups were treated equally except for addition of virus. All experiments resulted in blastocysts showing eGFP expression in both the trophoblast and the inner cell mass. In four experiments we used aliquots of the same virus preparation (1.25 � 107 IU/mL) for infection. The cleavage rate ranged from 50 to 70%, and blastocyst rate from 15 to 35%. In three experiments showing good to average embryo development, 25 to 83% of the resulting blastocysts expressed eGFP, indicating efficient lentiviral transduction of oocytes following laser-assisted zona microdrilling. Studies directly comparing zona microdrilling and subzonal injection with regard to the efficacy of lentiviral transgenesis and the incidence of polyspermy are underway.

Efficient transgenic rat production by a lentiviral vector

2007 ◽  
Vol 293 (1) ◽  
pp. H881-H894 ◽  
Author(s):  
Mieczyslaw Michalkiewicz ◽  
Teresa Michalkiewicz ◽  
Aron M. Geurts ◽  
Richard J. Roman ◽  
Glenn R. Slocum ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Lentiviral Vector ◽  
Insertion Site ◽  
Single Copy ◽  
Egfp Expression ◽  
Transgenic Rat ◽  
Sprague Dawley ◽  
Lentiviral Transgenesis ◽  
Sprague Dawley Rats ◽  
Transgenic Lines

A lentiviral construct for an enhanced green fluorescent protein (eGFP) driven by a chicken β-actin promoter, cytomegalovirus enhancer, and intronic sequences from rabbit β-globin (CAG) was used to produce transgenic lines of rats for evaluation of the usefulness of this approach in gene function studies. Fertilized eggs were collected from inbred Dahl S and outbred Sprague-Dawley rats, and ∼100 pl of concentrated virus were microinjected into the perivitrelline space of one-cell embryos. Of 121 embryos injected, 60 pups (49.6%) were born. Transgenic rates averaged 22% in Dahl S and 14% in Sprague-Dawley rats. Copy number ranged from one to four in the founders, and the inheritance of the transgene in a subsequent F1population was 48.2%. The small number of insertion sites enabled us to derive inbred transgenic lines with a single copy of the transgene within one generation. Sequencing of each transgene insertion site revealed that they inserted as single copies with a preference for the introns of genes. The CAG promoter drove high levels of eGFP expression in brain, kidney, heart, and vasculature, making it very suitable for exploring the cardiovascular function of newly discovered genes. The pattern of eGFP expression was similar across five different F1transgenic lines, indicating that the expression of the transgene was independent of its chromosomal position. Thus lentiviral transgenesis provides a powerful tool for the production of transgenic inbred rats and will enhance the usefulness of this species in gene discovery and target validation studies.

scholarly journals Lentiviral transgenesis of the leopard gecko, Eublepharis macularius

2014 ◽  
Vol 8 (10) ◽  
pp. 1070-1079 ◽  
Author(s):  
Hull Kaitlyn ◽  
Hodgson Dee ◽  
Clark Bob ◽  
W. Hickok Timothy ◽  
N. Petitte James ◽  
...  
Keyword(s):  
Eublepharis Macularius ◽  
Lentiviral Transgenesis ◽  
Leopard Gecko
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