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CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

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...

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Updated: Jun 18, 2025

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Engineering single-cycle MeV vector for CRISPR-Cas9 gene editing.

Ramya Rallabandi¹, Brenna Sharp², Spencer Majerus²

¹Mayo Clinic Graduate School of Biomedical Sciences, Virology and Gene Therapy Graduate Track, Mayo Clinic, Rochester, MN 55905, USA.

Molecular Therapy. Methods & Clinical Development

|July 29, 2024

Summary

This summary is machine-generated.

This study introduces a measles virus (MeV) vector for efficient CRISPR-Cas9 gene editing in human cells. The novel vector enables precise gene knock-out and knock-in, offering a flexible platform for genetic modification research.

Keywords:

CRISPR Cas9 HDR NHEJ gene editing iPSC measles viral vector

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Last Updated: Jun 18, 2025

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Published on: June 20, 2019

Area of Science:

Molecular Biology
Gene Editing Technologies
Virology

Background:

CRISPR-Cas9 is a powerful tool for gene editing with broad research and therapeutic potential.
Previous work established measles virus (MeV) as a single-cycle reprogramming vector for induced pluripotent stem cell generation.

Purpose of the Study:

To develop a single-cycle MeV vector for delivering CRISPR-Cas9 components (gRNA and Cas9 nuclease) into human cells.
To demonstrate the efficiency of this MeV vector for precise gene editing, including knock-out and knock-in.

Main Methods:

Construction of a single-cycle measles virus (MeV) vector encoding gRNA and Cas9 nuclease.
Delivery of the MeV vector to human cells for gene editing applications.
Assessment of on-target editing efficiency using reporter and endogenous genes (mCherry, HBB, FANCD1).
Evaluation of precise knock-in via homology-directed repair using oligonucleotide donors.

Main Results:

The MeV vector successfully mediated on-target gene editing in human cells.
Efficient editing was observed for both reporter (mCherry) and endogenous genes (HBB, FANCD1).
Precise gene knock-in was achieved using the MeV vector and a single-stranded oligonucleotide donor.

Conclusions:

The developed MeV vector serves as a novel and adaptable platform for gene knock-out and knock-in in human cells.
This system facilitates the integration of emerging gene editing technologies.
The MeV vector shows promise for advancing gene editing applications in basic and clinical research.