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In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

Genome Copying Errors

Genome Copying Errors

DNA replication is a well-evolved process that copies millions of base pairs with high fidelity during each cell division. Occasionally a wrong base or a long stretch of wrong bases may get added to the daughter strands. If the errors are left unchecked, cells might accumulate several mutations that might endanger their survival. Therefore, the copying errors are checked and repaired at three levels.

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Mutations in Microorganisms

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Related Experiment Video

Updated: Jan 1, 2026

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Published on: May 15, 2018

Efficient, continuous mutagenesis in human cells using a pseudo-random DNA editor.

Haiqi Chen¹, Sophia Liu^1,2,3, Samuel Padula¹

¹Broad Institute of MIT and Harvard, Cambridge, MA, USA.

Nature Biotechnology

|December 18, 2019

Summary

This summary is machine-generated.

Researchers developed a new gene editing tool called TRACE (T7 polymerase-driven continuous editing) for continuous, targeted mutagenesis in human cells. This method successfully identified mutations linked to MEK1 inhibitor resistance.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Published on: August 25, 2017

Area of Science:

Molecular Biology
Genetics
Cell Biology

Background:

Targeted mutagenesis is crucial for understanding gene function and disease mechanisms.
Existing methods for mutagenesis can be limited in efficiency, duration, or applicability in human cells.

Purpose of the Study:

To introduce a novel method for continuous, targeted mutagenesis in human cells.
To demonstrate the utility of this method in a drug resistance screening application.

Main Methods:

Developed TRACE (T7 polymerase-driven continuous editing), a system combining cytidine deaminase with T7 RNA polymerase.
Integrated a T7 promoter into the genome to drive continuous editing in specific genes.
Applied TRACE in a screen for MEK1 inhibitor resistance mutations.

Main Results:

TRACE enables high rates of mutagenesis over multiple cell generations.
The method successfully induced mutations in genes controlled by the T7 promoter.
Functionally correlated mutations conferring MEK1 inhibitor resistance were identified using TRACE.

Conclusions:

TRACE is an effective tool for continuous, targeted mutagenesis in human cells.
This method facilitates the discovery of functionally related mutations, particularly in drug resistance studies.
TRACE offers a powerful approach for genetic screens and functional genomics.