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Related Concept Videos

What is Gene Expression?01:36

What is Gene Expression?

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 processed and...
What is Gene Expression?01:42

What is Gene Expression?

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
A gene is a stretch of DNA that serves as the blueprint for functional RNAs and proteins. Since DNA is made up of nucleotides and proteins consist of amino...
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the addition of a...
Ribosome Profiling02:24

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique helps...
Leaky Scanning02:28

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
Structure of a Gene01:30

Structure of a Gene

A gene is the fundamental unit of heredity. Every individual has two copies of each gene, one inherited from each parent. Although most people contain the same genes, there is a small fraction that is slightly different amongst people. A gene with a small difference in its sequence of DNA bases forms different alleles, contributing to different phenotypes.
However, only 1% of the DNA is composed of genes that encode proteins; the rest, 99% is non-coding DNA. This non-coding DNA performs...

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Related Experiment Video

Updated: May 10, 2026

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

Tuning gene expression with synthetic upstream open reading frames.

Joshua P Ferreira1, K Wesley Overton, Clifford L Wang

  • 1Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA. cliff.wang@stanford.edu

Proceedings of the National Academy of Sciences of the United States of America
|June 27, 2013
PubMed
Summary

Researchers engineered short upstream open reading frames (uORFs) to precisely control recombinant protein expression levels. This novel method offers robust and predictable tuning of protein synthesis across various cell types.

Keywords:
Kozak consensus sequenceeukaryotic translationp21synthetic biologytranslation initiation site

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Related Experiment Videos

Last Updated: May 10, 2026

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters

Published on: November 26, 2013

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Gene Expression Regulation

Background:

  • Controlling recombinant protein expression levels is crucial for research and biotechnology.
  • Existing methods for tuning protein expression can be limited in range or predictability.

Purpose of the Study:

  • To develop a novel strategy for robust and tunable control of protein expression levels.
  • To engineer RNA sequence elements that can precisely modulate translation initiation.

Main Methods:

  • Engineered short upstream open reading frames (uORFs) encoding two-amino acid peptides.
  • Placed uORFs upstream of the protein of interest's coding sequence.
  • Varied sequences preceding uORFs to modulate translation initiation rates and suppression levels.

Main Results:

  • Achieved protein expression control spanning three orders of magnitude.
  • Demonstrated robust, predictable, and reproducible expression tuning across tested cell types.
  • Utilized multiple uORFs in series and non-AUG start codons for fine-tuning low expression levels.

Conclusions:

  • Engineered uORFs provide a versatile and effective method for controlling protein expression.
  • Translation initiation control is proposed as a primary strategy for tuning gene expression in mammalian systems.
  • This approach enables precise manipulation of protein synthesis for diverse applications.