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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.

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Related Experiment Video

Updated: Sep 29, 2025

Identification of Functionally-Relevant Lentivirus Integration Sites in an Insertional Mutagenesis Cell Library

Identification of Functionally-Relevant Lentivirus Integration Sites in an Insertional Mutagenesis Cell Library

Published on: January 10, 2025

Validation-Based Insertional Mutagenesis (VBIM), A Powerful Forward Genetic Screening Strategy.

Sarmishtha De¹, Ilaria Tamagno², George R Stark¹

¹Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio.

Current Protocols

|March 22, 2022

Summary

This summary is machine-generated.

Validation-Based Insertional Mutagenesis (VBIM) is a powerful forward genetics tool using lentiviruses for whole-genome screens. This method efficiently identifies genes influencing phenotypes through gain- or loss-of-function mutations.

Keywords:

cell libraries drug resistance forward genetics in vivo screens insertional mutagenesis lentiviral vectors

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Last Updated: Sep 29, 2025

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Published on: May 15, 2018

Area of Science:

Genetics
Molecular Biology
Genomics

Background:

Forward genetics identifies genes influencing phenotypes by analyzing genetic changes.
Traditional methods include cDNA, shRNA, CRISPR, and chemical mutagens, each with limitations.
Validation-Based Insertional Mutagenesis (VBIM) offers a novel approach using lentiviruses.

Purpose of the Study:

To review the VBIM strategy for genetic screens.
To highlight its application in identifying novel genes and functions.
To discuss its potential for in vivo applications.

Main Methods:

VBIM utilizes modified lentiviruses as insertional mutagens with strong promoters.
It generates large cell libraries with VBIM promoter insertions for phenotype-driven selection.
VBIM enables identification of gain- and loss-of-function mutations and targets any genomic locus.

Main Results:

VBIM facilitates selection of cells overexpressing VBIM-driven RNAs or proteins that promote a phenotype.
VBIM-driven RNAs can encode full-length, truncated, or antisense RNAs, impacting gene expression.
The strategy allows for true whole-genome screens without complex library construction.

Conclusions:

VBIM is a versatile and efficient strategy for forward genetic screens.
It has been successfully applied to identify novel genes and functions in diverse screens.
VBIM shows promise for transitioning to in vivo genetic screens.