Related Experiment Video

Updated: Jan 26, 2026

Using CRISPR/Cas9 Gene Editing to Investigate the Oncogenic Activity of Mutant Calreticulin in Cytokine Dependent Hematopoietic Cells
10:21

Using CRISPR/Cas9 Gene Editing to Investigate the Oncogenic Activity of Mutant Calreticulin in Cytokine Dependent Hematopoietic Cells

Published on: January 5, 2018

13.7K

Cell-specific CRISPR-Cas9 activation by microRNA-dependent expression of anti-CRISPR proteins

Mareike D Hoffmann1,2, Sabine Aschenbrenner1,2, Stefanie Grosse1

  • 1Synthetic Biology Group, Institute for Pharmacy and Biotechnology (IPMB) and Center for Quantitative Analysis of Molecular and Cellular Biosystems (BioQuant), University of Heidelberg, Heidelberg 69120, Germany.

Nucleic Acids Research
|April 16, 2019
PubMed

Insights

Researchers developed a novel CRISPR-Cas system that precisely targets gene editing in specific cells. This Cas-ON switch uses microRNAs to control anti-CRISPR proteins, enhancing safety for genetic therapies.

Area of Science:

  • Molecular Biology
  • Gene Editing Technologies
  • Biotechnology

Background:

  • CRISPR-Cas technologies offer promising avenues for personalized genetic disorder treatments.
  • Ensuring the precision and safety of CRISPR-based therapies necessitates strategies for localized Cas activity.
  • Confining Cas activity to specific cells and tissues is crucial for therapeutic applications.

Purpose of the Study:

  • To develop a cell type-specific Cas-ON switch for precise CRISPR-Cas activity control.
  • To engineer a system that restricts CRISPR-Cas9 function to targeted cells, enhancing therapeutic safety.
  • To create a modular platform adaptable to various CRISPR-Cas orthologues.

Main Methods:

  • Designed a Cas-ON switch utilizing miRNA-regulated expression of anti-CRISPR (Acr) proteins.
  • Inserted specific microRNA target sites (miR-122, miR-1) into Acr transgene 3'UTRs for cell-specific regulation.
  • Demonstrated functionality with Streptococcus pyogenes (Spy)Cas9 and Neisseria meningitidis (Nme)Cas9 orthologues.

Main Results:

  • Achieved cell-specific Cas9 activity in hepatocytes and cardiomyocytes via Acr knockdown.
  • Successfully inhibited Cas9 activity in off-target cells, ensuring safety.
  • Demonstrated control over genome editing and gene activation using the miR-dependent AcrIIA4 system.
  • Showcased system modularity by adapting it to NmeCas9 and its inhibitors.

Conclusions:

  • The developed Cas-ON switch enables precise, cell-specific CRISPR-Cas activity.
  • This system enhances the safety and efficacy of CRISPR-based gene therapies.
  • The modular design facilitates application with diverse CRISPR-Cas orthologues and anti-CRISPR proteins.

More Related Videos

Anti-RDL and Anti-mGlutR1 Receptors Antibody Testing in Honeybee Brain Sections using CRISPR-Cas909:25

Anti-RDL and Anti-mGlutR1 Receptors Antibody Testing in Honeybee Brain Sections using CRISPR-Cas9

Published on: January 30, 2020

8.6K
Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins10:46

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

2.3K

Related Concept Videos

CRISPR01:59

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...
57.6K
CRISPR and crRNAs02:53

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...
18.9K
CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
1.8K
Cell Specific Gene Expression01:58

Cell Specific Gene Expression

Multicellular organisms contain a variety of structurally and functionally distinct cell types, but the DNA in all the cells originated from the same parent cells. The differences in the cells can be attributed to the differential gene expression. Liver cells, whose functions include detoxification of blood, production of bile to metabolize fats, and synthesis of proteins essential for metabolism, must express a specific set of genes to perform their functions. Gene expression also varies with...
16.3K
Cell Specific Gene Expression01:58

Cell Specific Gene Expression

5.4K
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
24.1K