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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...

Crossing Over

Crossing Over

Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I, duplicated...

Crossing Over

Crossing Over

Unlike mitosis, meiosis aims for genetic diversity in its creation of haploid gametes. Dividing germ cells first begin this process in prophase I, where each chromosome—replicated in S phase—is now composed of two sister chromatids (identical copies) joined centrally.
The homologous pairs of sister chromosomes—one from the maternal and one from the paternal genome—then begin to align alongside each other lengthwise, matching corresponding DNA positions in a process called synapsis.
In order to...

Gene Conversion

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

Extraction: Partition and Distribution Coefficients

Extraction: Partition and Distribution Coefficients

The distribution law or Nernst's distribution law is the law that governs the distribution of a solute between two immiscible solvents. This law, also known as the partition law, states that if a solute is added to the mixture of two immiscible solvents at a constant temperature, the solute is distributed between the two solvents in such a way that the ratio of solute concentrations in the solvents remains constant at equilibrium.
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Separation of Sister Chromatids

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At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
At the onset of anaphase, separase, a proteolytic enzyme, is...

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Related Experiment Video

Updated: May 20, 2026

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

Published on: November 21, 2015

Intersectional Cre driver lines generated using split-intein mediated split-Cre reconstitution.

Ping Wang¹, Tianrui Chen, Katsuyasu Sakurai

¹Department of Cell Biology, Box 3709, Duke University Medical Center, Durham, NC, USA.

Scientific Reports

|July 10, 2012

Summary

This summary is machine-generated.

Scientists created a novel split-Cre system using split-inteins for precise gene targeting. This method enables highly specific Cre recombination in overlapping expression domains, advancing neuroscience research.

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Lineage Tracing and Clonal Analysis in Developing Cerebral Cortex Using Mosaic Analysis with Double Markers (MADM)

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Published on: May 8, 2020

Area of Science:

Neuroscience
Molecular Biology
Genetics

Background:

Developing tissue-specific Cre drivers for genetic research is challenging.
Existing Cre driver systems often lack the required specificity in complex tissues like the brain.

Purpose of the Study:

To develop a novel split-Cre system for achieving highly restricted Cre-recombination.
To enable intersectional targeting of gene expression in specific cell populations.

Main Methods:

Engineered a split-Cre system utilizing split-inteins for protein splicing-mediated Cre reconstitution.
Designed transgenic mice expressing split Cre components (Cre-N-intein-N and intein-C-Cre-C) driven by conserved human enhancers.
Validated Cre-dependent reporter gene induction in specific brain regions.

Main Results:

The split-intein-split-Cre system demonstrated highly efficient Cre reconstitution.
Cre-dependent reporter expression was specifically observed within the intersectional domains of the two driving enhancers.
The system successfully restricted Cre activity to the desired overlapping expression patterns.

Conclusions:

The split-intein-based split-Cre system offers a simpler and effective strategy for generating highly specific intersectional Cre drivers.
This method is particularly valuable for studying complex tissues, including the nervous system.
The system facilitates precise genetic manipulation in neuroscience research.