Transfection by Electroporation

Potter, Huntington; Heller, Richard

doi:10.1002/0471142301.nsa01es57

Cited by 9 publications

(6 citation statements)

References 33 publications

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“…The current study showed that comparing to the Bio-Rad electroporation buffer, the OptiMEM-GlutaMAX was compatible with iPS cells in terms of transfection rate and cell viability. However, unlike reported by Potter et al (15), conducting the electroporation in ice-cold medium did not improve the transfection efficiency in the current study, although the transfection efficiency of ice-cold medium was almost independent of pulsing condition.…”

Section: Discussioncontrasting

confidence: 99%

A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

Eghbalsaied

Hyder²,

Kues³

2019

Preprint

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A square-wave pulsing protocol was developed using OptiMEM-GlutaMAX for high efficient transfection of mouse embryonic fibroblast (MEF) and induced pluripotency stem (iPS) cells. An electrotransfection efficiency of > 95% was repeated for both MEF and iPS cells using reporter-encoding plasmids. The protocol was very efficient for plasmid size ranging from 6.2 to 13.5 kb. A high rate of targeted gene knockout (> 95 %) was produced in Venus transgenic cells using indels formation. Targeted deletions in the Venus transgene were performed by co-electroporation of two gRNA-encoding plasmids. In conclusion, this plasmid electrotransfection protocol is straight-forward, cost-effective, and efficient for CRISPRing mouse primary cells.

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Section: Discussioncontrasting

confidence: 99%

A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

Eghbalsaied

Hyder²,

Kues³

2019

Preprint

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“…marina is a naked dinoflagellate that had a difficult time withstanding the physical forces used in previously reported dinoflagellate transformation methods. Electroporation is a gentler method, allowing DNA to pass through temporary pores in an organism's membrane and has been utilized in many organisms, but requires the removal of seawater and replacement with electroporation buffers, often unable to maintain the osmolality of marine organisms 61 . A new electroporation model, Lonza's 4D-Nucleofector, provides a user with a score of built-in pulse settings and solutions that remove salts but help maintain dinoflagellates osmolality.…”

Section: Effectiveness Of Dna Introduction Methodsmentioning

confidence: 99%

Nuclear gene transformation in a dinoflagellate

Sprecher

Zhang

Lin

2019

Preprint

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The lack of a robust gene transformation tool that allows functional testing of the vast number of nuclear genes in dinoflagellates has greatly hampered our understanding of fundamental biology in this ecologically important and evolutionarily unique lineage. Here we report the development of a dinoflagellate expression vector, an electroporation protocol, and successful expression of introduced genes in the dinoflagellate Oxyrrhis marina. This protocol, involving the use of Lonza's Nucleofector and a codon optimized antibiotic resistance gene, has been successfully used to produce consistent results in several independent experiments. It is anticipated that this protocol will be adaptable for other dinoflagellates and will allow characterization of many novel dinoflagellate genes.

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“…However, in contrast to viral vectors with optimized strategies for cell transduction, nonviral vectors are still characterized by relatively low transfection efficiencies. Sonoporation, 14 electroporation (nucleofection) 15 and magnetofection 16 are interesting tools under development to enhance uptake of genetic material into cells.…”

Section: Introductionmentioning

confidence: 99%

Recent advances in gene‐enhanced bone tissue engineering

Betz

Kochanek

Rammelt

et al. 2018

The Journal of Gene Medicine

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The loss of bone tissue represents a critical clinical condition that is frequently faced by surgeons. Substantial progress has been made in the area of bone research, providing insight into the biology of bone under physiological and pathological conditions, as well as tools for the stimulation of bone regeneration. The present review discusses recent advances in the field of gene-enhanced bone tissue engineering. Gene transfer strategies have emerged as highly effective tissue engineering approaches for supporting the repair of the musculoskeletal system. By contrast to treatment with recombinant proteins, genetically engineered cells can release growth factors at the site of injury over extended periods of time. Of particular interest are the expedited technologies that can be applied during a single surgical procedure in a cost-effective manner, allowing translation from bench to bedside. Several promising methods based on the intra-operative genetic manipulation of autologous cells or tissue fragments have been developed in preclinical studies. Moreover, gene therapy for bone regeneration has entered the clinical stage with clinical trials for the repair of alveolar bone. Current trends in gene-enhanced bone engineering are also discussed with respect to the movement of the field towards expedited, translational approaches. It is possible that gene-enhanced bone tissue engineering will become a clinical reality within the next few years.

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Transfection by Electroporation

Cited by 9 publications

References 33 publications

A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

Nuclear gene transformation in a dinoflagellate

Recent advances in gene‐enhanced bone tissue engineering

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