Endpoint and Kinetic Approaches for Assessing Transfection Efficacy in Mammalian Cell Culture

• The efficacy of transfection reagents and nanoparticles is often assessed by measuring levels of expressed reporter protein. Fluorescence and luminescence based assays provide sensitive, quantifiable and repeatable approaches. The genes expressing reporter protein can be integrated into the cells to create stable reporter cell lines or can be expressed from a transfected plasmid. Green fluorescent protein, luciferase, and secreted alkaline phosphatase are well-established reporters with versatile applications. Monitoring changes in live cells during and after transfection offer opportunities to reveal related mechanisms, efficacy, and bottlenecks of transfection. In this chapter, we describe the experimental setup and considerations for in vitro screening of delivery vectors. This can further be extended to measurements in reporter cell lines.

Subject headings

NATURVETENSKAP  -- Biologi (hsv//swe)

NATURAL SCIENCES  -- Biological Sciences (hsv//eng)

MEDICIN OCH HÄLSOVETENSKAP  -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)

MEDICAL AND HEALTH SCIENCES  -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)

Keyword

Kinetic assay

Luciferase

Protein expression

Secreted alkaline phosphatase

Transfection

alkaline phosphatase

cell penetrating peptide

green fluorescent protein

plasmid vector

reagent

unclassified drug

adherent cell

cell culture

cell suspension

complex formation

cytolysis

flow cytometry

fluorescence

genetic transfection

in vitro study

incubation time

kinetics

luminescence

mammal cell

measurement

nonhuman

protein analysis

screening

animal

cell culture technique

cell line

genetics

mammal

plasmid

reporter gene

Animals

Cell Culture Techniques

Genes

Reporter

Mammals

Plasmids

Publication and Content Type

ref (subject category)

kap (subject category)