Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Cis-regulatory Sequences

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...

Cis-regulatory Sequences

Cis-regulatory Sequences

Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...

Introduction to Scalers

Introduction to Scalers

Many familiar physical quantities can be specified completely by giving a single number and the appropriate unit. For example, "a class period lasts 50 min," or "the gas tank in my car holds 65 L," or "the distance between the two posts is 100 m." A physical quantity that can be specified completely in this manner is called a scalar quantity. The word "scalar" is a synonym for "number." Time, mass, distance, length, volume, temperature, and energy are some examples of scalar quantities.
Scalar...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Strategy-based cognitive rehabilitation for childhood brain tumour: Protocol for an acceptability and feasibility trial of the Fatigue, Learning, and Memory Enrichment (FLaME) intervention.

NIHR open research·2025

Same author

The landscape of ketamine use disorder: Patient experiences and perspectives on current treatment options.

Addiction (Abingdon, England)·2025

Same author

Enabling participation in community-dwelling children and young people with acquired brain injuries and their families: a theory-based, evidence-based and person-based approach to intervention development.

BMJ open·2025

Same author

RNAi library screening reveals Gβ1, Casein Kinase 2 and ICAP-1 as novel regulators of LFA-1-mediated T cell polarity and migration.

Immunology and cell biology·2024

Same author

Evaluation of the quality and self-management related content of websites promoting physical activity engagement after myocardial infarction or cardiac surgery: A scoping review.

Heart & lung : the journal of critical care·2024

Same author

Understanding barriers and facilitators to long-term participation needs in children and young people following acquired brain injuries: a qualitative multi-stakeholder study.

Brain impairment : a multidisciplinary journal of the Australian Society for the Study of Brain Impairment·2024

Search research articles

Related Experiment Video

Updated: Jun 30, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

A Directed Evolution Protocol for Engineering Minimal Transcription Factors, Based on CIS Display.

Lin Qi¹, Emily Bennett¹, Mark Isalan²

¹Department of Life Sciences, Imperial College London, London, UK.

Methods in Molecular Biology (Clifton, N.J.)

|March 5, 2024

Summary

Directed evolution using cell-free DNA display (CIS display) enables high-throughput screening of DNA-binding proteins. This method overcomes limitations of traditional cloning, facilitating the evolution of synthetic biology components.

Keywords:

CIS display DNA-binding proteins Directed evolution High-throughput selection Protein engineering Transcription factors (TF)

More Related Videos

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

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

Published on: November 26, 2013

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

Related Experiment Videos

Last Updated: Jun 30, 2026

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Published on: March 16, 2011

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

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

Published on: November 26, 2013

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening

Published on: April 1, 2016

Area of Science:

Molecular Biology
Synthetic Biology
Biotechnology

Background:

Directed evolution is key for creating novel biomolecules, with display techniques enabling high-throughput screening.
Classical methods using plasmid cloning limit library sizes (~10^6-10^7) due to transformation efficiencies.
In vitro display methods, like CIS display, bypass cloning, enabling vastly larger library sizes (>10^12).

Purpose of the Study:

To adapt the cell-free DNA display (CIS display) technique for the high-throughput selection of DNA-binding proteins.
To demonstrate the efficacy of CIS display for evolving functional DNA-binding proteins, a class not previously targeted by this method.
To provide a protocol applicable to engineering transcription factors and other DNA-binding proteins for synthetic biology.

Main Methods:

Utilized cell-free DNA display (CIS display), a DNA-based in vitro technique.
Leveraged the RepA DNA replication initiator protein to link genotype to phenotype.
Applied the method to select functional DNA-binding proteins from large combinatorial libraries.

Main Results:

Successfully adapted CIS display for the selection of DNA-binding proteins.
Demonstrated efficient enrichment of the minimal transcription factor Cro from a low starting frequency (1 in 10^9).
Validated the protocol's applicability for evolving DNA-binding proteins and transcription factors.

Conclusions:

CIS display is an effective method for the directed evolution of DNA-binding proteins.
This technique overcomes previous library size limitations, expanding possibilities in protein engineering.
The protocol is valuable for advancing mammalian synthetic biology by enabling the engineering of crucial DNA-binding components.