Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Mutations in Microorganisms01:18

Mutations in Microorganisms

44
Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
44
Mismatch Repair01:20

Mismatch Repair

5.0K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
5.0K
Point and Frameshift Mutations01:30

Point and Frameshift Mutations

53
Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...
53
Mutations01:39

Mutations

84.0K
Overview
84.0K
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

87
Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
87

You might also read

Related Articles

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

Sort by
Same author

Anti-CRISPR-mediated continuous directed evolution of CRISPR-Cas9 in human cells.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Optimizing Radiography Utilization: Multidisciplinary Expert Consensus Recommendations Endorsed by the Society of Academic Bone Radiologists, Society of Skeletal Radiology, American Society of Emergency Radiology, Orthopaedic Trauma Association, American Academy of Emergency Medicine, and American Rhinologic Society.

Radiology·2026
Same author

IMPACT OF FLUORESCENT DYES ON MUTATIONS IN NEXT GENERATION SEQUENCING LIRBARY PREPARATION.

bioRxiv : the preprint server for biology·2026
Same author

Hybrid capture RNA-seq defines temporal gene expression in <i>Rickettsia</i>.

mSphere·2026
Same author

Brief Report: Androgen Receptor Expression Is Associated With Male Predominance in Thymic Neuroendocrine Neoplasms.

JTO clinical and research reports·2026
Same author

Assessing Internist Competency in Point-of-Care Ultrasound Using the Entrustable Professional Activity Framework.

Journal of general internal medicine·2026

Related Experiment Video

Updated: Aug 12, 2025

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

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

Published on: March 16, 2011

30.6K

Expanded MutaT7 toolkit efficiently and simultaneously accesses all possible transition mutations in bacteria.

Amanuella A Mengiste1, Robert H Wilson1, Rachel F Weissman1

  • 1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.

Nucleic Acids Research
|January 30, 2023
PubMed
Summary
This summary is machine-generated.

Scientists developed a more active enzyme fusion (eMutaT7A→G) for faster genetic diversification. This tool, combined with another enzyme, can now create all transition mutations simultaneously for directed evolution in bacteria.

More Related Videos

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
11:36

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Published on: July 3, 2016

11.0K
Following the Dynamics of Structural Variants in Experimentally Evolved Populations
04:52

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Published on: February 3, 2023

1.0K

Related Experiment Videos

Last Updated: Aug 12, 2025

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli
09:01

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

Published on: March 16, 2011

30.6K
A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
11:36

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Published on: July 3, 2016

11.0K
Following the Dynamics of Structural Variants in Experimentally Evolved Populations
04:52

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Published on: February 3, 2023

1.0K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetics

Background:

  • Nucleotide base deaminase-T7 RNA polymerase fusions are effective for in vivo directed evolution.
  • Existing tools enable genetic diversification but can be improved for efficiency.

Purpose of the Study:

  • To enhance the utility of deaminase-T7 RNA polymerase fusions for directed evolution.
  • To introduce a highly active adenosine deaminase-T7 RNA polymerase fusion (eMutaT7A→G) for increased mutation frequencies.
  • To demonstrate the simultaneous induction of all transition mutations using combined adenosine and cytidine deaminase fusions.

Main Methods:

  • Development and characterization of an enhanced adenosine deaminase-T7 RNA polymerase fusion (eMutaT7A→G).
  • Assessment of mutation frequencies and potential benefits/downsides of the enhanced mutator in Escherichia coli.
  • Co-expression of adenosine deaminase-bearing mutators (MutaT7A→G or eMutaT7A→G) with a cytidine deaminase-bearing mutator (MutaT7C→T).

Main Results:

  • The eMutaT7A→G fusion significantly increased mutation frequencies, enabling more rapid directed evolution.
  • Adenosine and cytidine deaminase mutators were successfully used in tandem to induce all transition mutations simultaneously in E. coli.
  • The approach was validated by exploring pathways to antibacterial drug resistance.

Conclusions:

  • Optimized MutaT7 tools, including the highly active eMutaT7A→G, accelerate in vivo directed evolution.
  • The ability to induce all transition mutations simultaneously broadens the scope of directed evolution applications.
  • This strategy is poised for broad application across diverse organisms for genetic diversification and trait optimization.