Non-viral Expression
Non-viral Expression - Related Videos
10:48
Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
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Cited by 5 •
2014
We describe the usage of a fluorescent reporter vaccinia virus that enables real-time measurement of viral infectivity and gene expression through the stage-specific expression of spectrally distinct reporter fluorophores. We detail a plate-based method for accurately identifying the stage at which virus replication is affected in response to small molecule inhibition.
05:28
Targeted Expression of GFP in the Hair Follicle Using Ex Vivo Viral Transduction
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Cited by 1 •
2008
The hair follicle is a highly complex appendage of the skin containing a multiplicity of cell types. The follicle undergoes constant cycling through the life of the organism including growth and resorption with growth dependent on specific stem cells. The targeting of the follicle by genes and stem cells to change its properties, in particular, the nature of the hair shaft is, discussed. Hair follicle delivery systems are described, such as liposomes and viral vectors for gene therapy. The...
11:08
Packaging HIV- or FIV-based Lentivector Expression Constructs & Transduction of VSV-G Pseudotyped Viral Particles
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Cited by 15 •
2012
Lentiviral expression vectors are the most effective vehicles for stably expressing different effector molecules or reporter constructs in dividing and non-dividing mammalian cells and whole organisms. Here we provide a protocol on how to package lentivector expression constructs in pseudoviral particles and to transduce target cells using the pseudoviral particles.
08:22
Stereotaxic Microinjection of Viral Vectors Expressing Cre Recombinase to Study the Role of Target Genes in Cocaine Conditioned Place Preference
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Cited by 2 •
2013
This article describes how to microinject viral vectors into mouse brain and then test in a conditioned place preference paradigm that includes an acquisition, extinction and reinstatement phase.
08:26
Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney
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Cited by 10 •
2018
This protocol describes a method to inject plasmid DNA into the mouse kidney via the renal pelvis to produce transgene expression specifically in the kidney.
FAQs
Expression depends on where the introduced cargo becomes available to cellular machinery and on how long it remains functional. Plasmid DNA is associated with delivery toward the nucleus, whereas messenger RNA acts in the cytoplasm, so the intended destination can influence onset and persistence. This distinction helps researchers select a delivery strategy for a particular cancer experiment.
Chemical transfection, electroporation, and lipid- or polymer-based carriers provide different ways to move nucleic acids across the cell membrane. Their importance is practical: the delivery format must be matched to the cargo and cell system so that expression is sufficient without excessive loss of viability. Comparing these options lets investigators tune the balance rather than treating delivery as a fixed step.
Three outcomes are especially important when optimizing a non-viral system: expression level, duration, and cell viability. Increasing one outcome may not produce a useful experiment if the cells are damaged or expression disappears too quickly. Cancer researchers therefore adjust delivery conditions and evaluate these measures together, creating a system suited to the biological question rather than maximizing protein production alone.
A typical experimental workflow begins by choosing the genetic cargo, such as plasmid DNA or messenger RNA, and then selecting chemical transfection, electroporation, or a lipid- or polymer-based carrier. After delivery, investigators assess whether the intended protein or functional RNA is produced and whether cells remain viable. These readouts guide further adjustment of the conditions.
Non-viral expression supports tests of oncogene and tumor-suppressor function by allowing researchers to introduce genetic material and observe its resulting activity in cells. It can also support evaluation of gene-silencing strategies. These applications help connect a candidate gene or nucleic-acid intervention with cellular behavior during cancer research, before investigators refine delivery conditions for subsequent studies.
Engineered immune cells are one important application in cancer research. Non-viral delivery can provide genetic material to these cells for expression studies or cell engineering, while the same general approach can be used in cancer-vaccine development and other targeted treatments. The relevant design question is whether the chosen cargo and delivery conditions produce the required expression while preserving cell viability.