Expression Constructs
Expression Constructs - Related Videos
08:25
Phage-mediated Delivery of Targeted sRNA Constructs to Knock Down Gene Expression in E. coli
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Cited by 5 •
2016
We describe a method to knock down gene expression in a growing population of E. coli cells using sequence-targeted sRNA expression cassettes delivered by an M13 phagemid vector.
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.
10:05
Improved Generation of Induced Cardiomyocytes Using a Polycistronic Construct Expressing Optimal Ratio of Gata4, Mef2c and Tbx5
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Cited by 35 •
2015
We describe here a protocol for the generation of iCMs using retrovirus-mediated delivery of Gata4, Tbx5 and Mef2c in a polycistronic construct. This protocol yields a relatively homogeneous population of reprogrammed cells with improved efficiency and quality and is valuable for future studies of iCM reprogramming.
07:30
HSV-Mediated Transgene Expression of Chimeric Constructs to Study Behavioral Function of GPCR Heteromers in Mice
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Cited by 8 •
2016
This article describes how to inject viral vectors into the mouse frontal cortex to test behavioral assays that require GPCR heteromeric formation.
09:59
AAV Deployment of Enhancer-Based Expression Constructs In Vivo in Mouse Brain
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Cited by 3 •
2022
This protocol describes a novel rAAV-based transient enhancer-reporter assay. This assay can be used to induce enhancer-driven expression in vivo in the mouse brain.
FAQs
Promoters initiate transcription, while regulatory elements help control how strongly and consistently the selected coding sequence is transcribed. These choices can therefore alter the amount of messenger RNA available for translation and, ultimately, the quantity of gene product produced. In immune and infection experiments, this control helps researchers obtain expression levels appropriate for functional assays, diagnostics, or mechanistic studies.
Changes to an expression construct can affect more than total production. Construct design may influence the gene product’s cellular localization and protein stability, which determine whether the product is present in the relevant cellular compartment and remains suitable for analysis. These properties matter when researchers evaluate recombinant antigens, antibodies, cytokines, or pathogen proteins in controlled experiments.
The host cell supplies the biological setting in which the construct is transcribed into messenger RNA and the resulting message is translated into protein. Consequently, expression is not determined by the DNA sequence alone. The relationship between construct design and host-cell activity affects the amount and usable form of the product available for immunology or infection research.
The promoter and regulatory elements first govern transcription of the coding sequence into messenger RNA. That RNA then serves as the template for translation, producing the selected protein or other gene product. This two-stage pathway explains why design decisions that change transcription can influence downstream protein production and the reproducibility of assays using the expressed material.
Planning should specify the desired gene product, the coding sequence that will represent it, and the promoter and regulatory elements needed to control its expression. Researchers should also consider the intended host cell and the product’s required localization and stability. Defining these features in advance links construct architecture to the planned experimental readout.
Researchers use these constructs to produce recombinant antigens, antibodies, cytokines, and pathogen proteins. The resulting products support functional assays that examine biological activity, diagnostic studies that evaluate detection, vaccine studies that investigate immune-relevant materials, and mechanistic experiments designed to clarify interactions between immune responses and infectious agents.
They allow investigators to connect a selected gene product with measurable immune or infection-related outcomes. Depending on the product and experiment, researchers can assess function, support diagnostic development, investigate vaccine-related questions, or examine mechanisms involving immune responses and pathogen proteins. Expression level, localization, and stability also help determine how reliably those outcomes can be interpreted.