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Related Concept Videos

DNA-only Transposons02:57

DNA-only Transposons

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DNA-only transposons are called autonomous transposons since they code for the enzyme transposase that is required for the transposition mechanism. Insertion of transposons can alter gene functions in multiple ways. They can mutate the gene, alter gene expression by introducing a novel promoter or insulator sequence, introduce new splice sites, and change the mRNA transcripts produced, or remodel chromatin structure.
The donor site from where the transposon is excised is either degraded or...
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What is Gene Expression?01:42

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
Genetic Information Flows from DNA to RNA to Protein
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What is Gene Expression?01:36

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A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is comprised  of nucleotides and proteins are comprised of amino acids, a mediator is required to convert the information encoded in DNA into proteins. This mediator is the messenger RNA (mRNA). mRNA copies the blueprint from DNA by a process called transcription. In eukaryotes, transcription occurs in the nucleus by complementary base-pairing with the DNA template. The mRNA is then...
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Cell Specific Gene Expression01:58

Cell Specific Gene Expression

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Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
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Chromatin Position Affects Gene Expression02:35

Chromatin Position Affects Gene Expression

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Chromatin is the massive complex of DNA and proteins packaged inside the nucleus. The complexity of chromatin folding and how it is packaged inside the nucleus greatly influences  access to genetic information. Generally, the nucleus' periphery is considered transcriptionally repressive, while the cell's interior is considered a transcriptionally active area. 
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Related Experiment Video

Updated: Feb 1, 2026

Electroporation-Based Genetic Modification of Primary Human Pigment Epithelial Cells Using the Sleeping Beauty Transposon System
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High-performance gene expression and knockout tools using sleeping beauty transposon system.

Kaishun Hu1, Yu Li1, Wenjing Wu1,2

  • 11Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, 510120 China.

Mobile DNA
|December 12, 2018
PubMed
Summary

New Sleeping Beauty (SB) transposon vectors enable efficient gene overexpression and knockout in mammalian cells. These pSB vectors facilitate protein purification and tracking, advancing molecular cell biology research.

Keywords:
BRD7CRISPRFBW7NFATc1RCC2Sleeping beauty

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Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • DNA transposons, like the Sleeping Beauty (SB) system, offer stable gene expression in mammalian cells.
  • Existing SB transposon systems lack tools for protein purification/tracking and gene knockout.

Purpose of the Study:

  • To develop novel SB-derived vectors (pSB vectors) for versatile gene manipulation in mammalian cells.
  • To create tools for protein purification, tracking, and gene knockout using the SB transposon system.

Main Methods:

  • Generated pSB vectors incorporating Sleeping Beauty inverted repeat-direct repeat-left/right (IRDR-L/R) sequences for transposition by SB transposase.
  • Integrated Gateway cassettes for simplified cloning and various tags (Flag, Myc, HA, V5, SFB) for protein detection.
  • Incorporated CRISPR-Cas9 cassette into pSB plasmids for targeted gene knockout.

Main Results:

  • Successfully generated a series of pSB vectors suitable for gene cloning and expression.
  • Utilized pSB-SFB-GFP vector for Tandem Affinity Purification, identifying NFATc1 as a novel FBW7 binding partner.
  • Employed pSB-CRISPR vectors to knock out RCC2 and BRD7, revealing their roles in mitosis.

Conclusions:

  • The pSB vector series provides a convenient and powerful platform for gene overexpression and knockout in mammalian cells.
  • These vectors represent a valuable new alternative for molecular and cell biology research.