ArticleProceedings of the National Academy of Sciences of the United States of America1987
Transfection of mammalian cells with plasmid DNA by scrape loading and sonication loading.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1987. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 73 papers.
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Who cites it
73 citing papers in PubMed, 311 citations in OpenAlex.
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- Microfluidic and Nanofluidic Intracellular Delivery.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2021Review
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- Acoustofluidic sonoporation for gene delivery to human hematopoietic stem and progenitor cells.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Instant labeling of therapeutic cells for multimodality imaging.Theranostics · 2020Article
- Nano-Enhanced Drug Delivery and Therapeutic Ultrasound for Cancer Treatment and Beyond.Frontiers in bioengineering and biotechnology · 2019Review
- Targeting silencing androgen receptor gene by shRNA with low-intensity focused ultrasonic irradiation inhibits growth of prostate cancer xenografts in nude mice.International journal of clinical and experimental pathology · 2019Article
- Oncolytic herpes simplex virus and immunotherapy.BMC immunology · 2018Review
13 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
Scrape loading and sonication loading are two recently described methods of introducing macromolecules into living cells. We have tested the efficacy of these methods for transfection of mammalian cells with exogenous DNA, using selection systems based either on resistance to the drug G418 (Geneticin) or on acquisition of the ability to utilize the salvage pathway of pyrimidine biosynthesis. These loading methods can be employed to generate cell lines that express the gene product of the transfected DNA molecules both transiently and stably. Optimal transfection is observed when the DNA is added to cells in physiological saline lacking divalent cations and containing K+ in place of Na+. DNA molecules 7.1 to 30 kilobases long have been introduced by the scrape loading procedure. In addition, the scrape loading procedure has been employed for cotransfection and subsequent expression of nonselectable genes encoded on DNA molecules added in a mixture with DNA molecules whose expression is selected. Cell lines expressing oncogenes or proteins that are important for regulation of cell growth and division have been obtained by this procedure. The scrape loading procedure is also useful for studies of the cellular changes that occur upon expression of an exogenous gene. As many as 80% of cells scrape loaded with the plasmid pC6, which encodes the simian virus 40 large tumor antigen, contained this protein in the nucleus between 1 and 5 days after transfection. Thus, scrape loading and sonication loading are simple, economical, and reproducible methods for introduction of DNA molecules into adherent and nonadherent cells, and these methods may be useful in the future for experimentation at both fundamental and applied levels.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.