Transgenesis Protocol
Transgenesis Protocol - Related Videos
07:23
Mosaic Zebrafish Transgenesis for Evaluating Enhancer Sequences
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Cited by 6 •
2010
We demonstrate our approach to finding potential enhancer elements from developmentally regulated genes and evaluating their function through mosaic zebrafish transgenesis.
08:51
Microinjection for Transgenesis and Genome Editing in Threespine Sticklebacks
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Cited by 30 •
2016
Transgenic manipulations and genome editing are critical for functionally testing the roles of genes and cis-regulatory elements. Here a detailed microinjection protocol for the generation of genomic modifications (including Tol2-mediated fluorescent reporter transgene constructs, TALENs, and CRISPRs) is presented for the emergent model fish, the threespine stickleback.
06:57
RNAi Mediated Gene Knockdown and Transgenesis by Microinjection in the Necromenic Nematode Pristionchus pacificus
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Cited by 28 •
2011
In model organisms, transgenesis can manipulate gene functions while RNAi can knockdown specific mRNA transcripts 1-2. This protocol aims to illustrate the techniques needed to introduce stably transmitted DNA and transient double stranded RNA into the necromenic nematode Pristionchus pacificus for studies in evolutionary, developmental, and behavioral biology.
05:32
Embryo Microinjection for Transgenesis in Drosophila
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2024
This article takes the phiC31 integrase-mediated transgenesis in Drosophila as an example and presents an optimized protocol for embryo microinjection, a crucial step for creating transgenic flies.
05:06
An Efficient Transgenesis Approach for Gene Delivery in the Mouse Embryonic Heart
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2024
This protocol presents a detailed methodological framework for electroporation-based transgenesis of cardiac cells in developing mouse hearts. The video assets provided here will facilitate learning of this versatile technique.
FAQs
The DNA construct determines which foreign gene is introduced and provides the material whose integration and expression will later be evaluated. Its design therefore connects the experimental question to the resulting genetic change. A carefully planned construct helps researchers investigate gene function, regulation, or effects on development and physiology after the modified organism is examined.
Delivery into cells or early embryos creates an opportunity for the introduced DNA to become part of the developing organism. When integration occurs at an appropriate stage, researchers can identify individuals carrying the transgene and assess whether the change is maintained through development. This approach supports studies of organism-level traits rather than isolated cellular effects.
These outcomes describe different stages of validation. Integration asks whether the foreign DNA entered the genome, expression asks whether the added gene is active, and inheritance asks whether the change passes to subsequent generations. Separating these questions prevents the presence of a transgene from being treated as proof that it functions or is heritable.
Molecular analyses provide evidence that the intended genetic change occurred and help assess its biological status. Researchers use them to confirm integration, examine expression, and evaluate inheritance. These results establish whether an individual is suitable for downstream studies, including analyses of development, physiology, or disease-related traits associated with the introduced gene.
The workflow begins with designing a DNA construct, followed by delivering it into cells or early embryos. Researchers then identify individuals in which the transgene has integrated and is expressed. Molecular confirmation of integration and inheritance comes before examining effects on development, physiology, disease-related traits, or other characteristics relevant to the study.
Transgenesis is useful when researchers need to connect a specific gene with a biological function or trait. It supports functional gene studies, investigation of gene regulation, and creation of animal models for disease-related research. The same general strategy can also produce organisms with useful research or biotechnological characteristics, extending its value beyond basic biology.