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

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

You might also read

Related Articles

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

Sort by

Same author

CNR1 and CNR2 Cannabinoid Receptor Mutations in Cancer Cells.

Current issues in molecular biology·2026

Same author

Cancer mutations in RAD51 and its paralogues.

PloS one·2026

Same author

MS-ConTab: multi-scale contrastive learning of mutation signatures for Pan-Cancer representation and stratification.

Bioinformatics (Oxford, England)·2026

Same author

Nucleotide Substitution Biases in Related Cancer Driver Genes.

International journal of molecular sciences·2025

Same author

Mutations altering the DNA binding domains of the human RAD52 protein exert distinct effects on homologous recombination repair in Saccharomyces cerevisiae.

G3 (Bethesda, Md.)·2025

Same author

PRMT5 genetic interactions with DNA double strand break repair genes.

PloS one·2025

Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026

Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Jun 7, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

SIMPLE Method for Plasmid Editing.

Ruben C Petreaca^1,2, Hovik J Gasparyan³

¹Department of Molecular Genetics, The Ohio State University, Marion, OH, USA.

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

|November 11, 2024

Summary

This summary is machine-generated.

Molecular cloning techniques have advanced gene function studies. A new method, SIMPLE (SapI/AarI incision-mediated plasmid editing), enables precise plasmid modifications like deletions and insertions.

Keywords:

Molecular cloning Plasmid editing Site-directed mutagenesis

More Related Videos

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

Homemade Site Directed Mutagenesis of Whole Plasmids

Homemade Site Directed Mutagenesis of Whole Plasmids

Published on: May 11, 2009

Related Experiment Videos

Last Updated: Jun 7, 2025

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

Homemade Site Directed Mutagenesis of Whole Plasmids

Homemade Site Directed Mutagenesis of Whole Plasmids

Published on: May 11, 2009

Area of Science:

Molecular Biology
Genetic Engineering

Background:

Molecular cloning techniques, including PCR and site-specific mutagenesis, have been crucial for understanding gene function.
These methods are widely used in biological research laboratories.

Purpose of the Study:

To introduce and describe a novel molecular cloning technique called SIMPLE (SapI/AarI incision-mediated plasmid editing).
To provide detailed instructions for implementing the SIMPLE cloning method.

Main Methods:

The study details a new technique, SIMPLE cloning, which utilizes SapI/AarI restriction enzymes for plasmid editing.
This method facilitates efficient deletions, short insertions (e.g., epitope tagging), and accurate mutations in episomal plasmids.

Main Results:

SIMPLE cloning offers a precise and efficient way to modify episomal plasmids.
The technique is applicable for various genetic manipulations, including deletions, insertions, and mutations.

Conclusions:

SIMPLE cloning represents a valuable addition to the molecular geneticist's toolkit.
This method enhances the precision and efficiency of plasmid engineering for biological research.