Related Experiment Video

Updated: Dec 17, 2025

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays
14:06

Mapping Bacterial Functional Networks and Pathways in Escherichia Coli using Synthetic Genetic Arrays

Published on: November 12, 2012

46.9K

Identification and Characterization of a Neutral Locus for Knock-in Purposes in C. parapsilosis

Tibor Nemeth1, Csaba Papp1, Csaba Vagvolgyi1

  • 1Department of Microbiology, University of Szeged, Szeged, Hungary.

Frontiers in Microbiology
|June 26, 2020
PubMed

Insights

Researchers identified a new genetic locus, CpNEUT5L (N5L), in Candida parapsilosis. This neutral site enables efficient gene knock-in, aiding the study of fungal pathogens and their virulence factors.

Area of Science:

  • Medical Mycology
  • Molecular Biology
  • Genetics

Background:

  • Invasive fungal infections caused by Candida species are a significant global health concern, with Candida parapsilosis posing a particular threat to neonates.
  • Investigating Candida parapsilosis virulence requires genetic manipulation tools, but suitable knock-in loci have been lacking.

Purpose of the Study:

  • To identify and characterize a novel, neutral genetic locus in Candida parapsilosis for gene knock-in applications.
  • To develop and validate plasmids for targeting this locus using both Gateway and classical ligation methods.

Main Methods:

  • Identification of the intergenic locus CpNEUT5L (N5L) in Candida parapsilosis.
  • Development of integrative replacement plasmids for Gateway and classical ligation.
  • Fluorescent tagging of laboratory and prototroph strains by integrating constructs into the N5L locus.
  • Assessment of growth under stress conditions and basic virulence properties of modified strains.

Main Results:

  • The CpNEUT5L (N5L) locus was successfully utilized for gene knock-in in Candida parapsilosis.
  • Integration of GFP-expressing constructs yielded strong, homogenous fluorescent signals.
  • Mutants with alterations at the N5L locus exhibited no significant changes in growth or basic virulence properties, even under stress.

Conclusions:

  • CpNEUT5L (N5L) is a highly effective neutral locus for knock-in approaches in Candida parapsilosis.
  • This locus facilitates the study of fungal pathogen virulence and the development of new therapeutic strategies.
Keywords:
Candida parapsilosisGFPgenetic manipulationknock-inoverexpressionreintegration

More Related Videos

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines11:32

Gene Knock-in by CRISPR/Cas9 and Cell Sorting in Macrophage and T Cell Lines

Published on: November 13, 2021

11.3K
A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins08:37

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

8.0K

Related Concept Videos

Conservative Site-specific Recombination and Phase Variation02:53

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...
6.5K
In-vitro Mutagenesis01:16

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.
15.9K