Electroporation: Past, present and future

Nakamura, Harukazu; Funahashi, Junichi

doi:10.1111/dgd.12012

Cited by 45 publications

(30 citation statements)

References 58 publications

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“…This approach creates genetic mutations in a subset of cells within a wild-type background, a technology that was used extensively in the Drosophila field to study complex biological processes (Blair, 2003). The electroporation technique, extensively used in the chicken embryo (Itasaki et al, 1999;Nakamura and Funahashi, 2013;Scaal et al, 2004;Serralbo et al, 2013;Voiculescu et al, 2008;Yokota et al, 2011), also results in the mosaic expression of constructs, which, combined to lossof-function approaches, could provide similar advantages as in fly. However, gene inactivation in the chicken has been limited to knockdown by RNA interference-and morpholino-based methodologies (Das et al, 2006;Gros et al, 2009;Hou et al, 2011;Norris and Streit, 2014;Rios et al, 2011;Serralbo and Marcelle, 2014;Voiculescu et al, 2008) that each have their own limitations, including variability in the level of knockdown, off target effects, and transient inhibition of transcripts.…”

Section: Introductionmentioning

confidence: 99%

CRISPR mediated somatic cell genome engineering in the chicken

Véron

Kipen

et al. 2015

Developmental Biology

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Gene-targeted knockout technologies are invaluable tools for understanding the functions of genes in vivo. CRISPR/Cas9 system of RNA-guided genome editing is revolutionizing genetics research in a wide spectrum of organisms. Here, we combined CRISPR with in vivo electroporation in the chicken embryo to efficiently target the transcription factor PAX7 in tissues of the developing embryo. This approach generated mosaic genetic mutations within a wild-type cellular background. This series of proof-of-principle experiments indicate that in vivo CRISPR-mediated cell genome engineering is an effective method to achieve gene loss-of-function in the tissues of the chicken embryo and it completes the growing genetic toolbox to study the molecular mechanisms regulating development in this important animal model.

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Section: Introductionmentioning

confidence: 99%

CRISPR mediated somatic cell genome engineering in the chicken

Véron

Kipen

et al. 2015

Developmental Biology

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“…It ranges from values of approximately 100 V/cm in large cells up to 1-2 kV/cm in small cells such as bacteria. Plasmid electrotransfer is a multistep process from interaction with the cell membrane, movement into the cell, intracellular trafficking and passage across the nuclear membrane [Wells, 2010;Nakamura and Funahashi, 2013]. A variety of different electrodes could be used depending on the cells to be treated.…”

Section: Applications Of Electroporationmentioning

confidence: 99%

Electroporation – Advantages and Drawbacks for Delivery of Drug, Gene and Vaccine

Bolhassani¹,

Khavari²,

Orafa³

2014

Application of Nanotechnology in Drug Delivery

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“…This allows us to perform lineage studies to follow the fate and movements of cells during development but also perform gain and loss of function of genes of interest. This can be coupled with the in vivo observation of cell behavior, using classical and two photon confocal microscope technologies (Itasaki et al 1999;Nakamura et al 2004;Uchikawa 2008;Chuai et al 2009;Yokota et al 2011;Rios et al 2012;Nakamura and Funahashi 2013;Serralbo et al 2013;Kulesa et al 2013). The combination of these technologies opens new fields of investigation, until now restricted to more simple systems and makes the chick embryo one of the most exciting and versatile model to characterize in an amniote environment dynamic processes, such as tissue morphogenesis, cell migration and proliferation, signaling, etc.…”

Section: Head and Limb Musclesmentioning

confidence: 99%

The Avian Embryo as a Model System for Skeletal Myogenesis

Hirst

Marcelle

2014

Results and Problems in Cell Differentiation

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This review will focus on the use of the chicken and quail as model systems to analyze myogenesis and as such will emphasize the experimental approaches that are strongest in these systems-the amenability of the avian embryo to manipulation and in ovo observation. During somite differentiation, a wide spectrum of developmental processes occur such as cellular differentiation, migration, and fusion. Cell lineage studies combined with recent advancements in cell imaging allow these biological phenomena to be readily observed and hypotheses tested extremely rapidly-a strength that is restricted to the avian system. A clear weakness of the chicken in the past has been genetic approaches to modulate gene function. Recent advances in the electroporation of expression vectors, siRNA constructs, and use of tissue specific reporters have opened the door to increasingly sophisticated experiments that address questions of interest not only to the somite/muscle field in particular but also fundamental to biology in general. Importantly, an ever-growing body of evidence indicates that somite differentiation in birds is indistinguishable to that of mammals; therefore, these avian studies complement the complex genetic models of the mouse.

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Electroporation: Past, present and future

Cited by 45 publications

References 58 publications

CRISPR mediated somatic cell genome engineering in the chicken

CRISPR mediated somatic cell genome engineering in the chicken

Electroporation – Advantages and Drawbacks for Delivery of Drug, Gene and Vaccine

The Avian Embryo as a Model System for Skeletal Myogenesis

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