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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...

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

CRISPR/Cas9 Genome Editing

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...

CRISPR

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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...

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Calcium-based nanomaterials and their interrelation with chitosan: optimization for pCRISPR delivery.

Navid Rabiee¹, Mojtaba Bagherzadeh¹, Amir Mohammad Ghadiri¹

¹Department of Chemistry, Sharif University of Technology, Tehran, Iran.

Journal of Nanostructure in Chemistry

|September 28, 2021

Summary

Calcium and chitosan blends show promise for delivering CRISPR gene-editing tools. These novel nanoparticles enhance gene delivery efficiency in human cells, offering a potential advancement in treating genetic diseases.

Keywords:

Calcium-based non-viral vector Chitosan-based nanomaterials Gene delivery pCRISPR

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Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Published on: March 25, 2015

Area of Science:

Biomaterials Science
Nanotechnology
Gene Therapy

Background:

CRISPR technology offers potential for treating genetic diseases.
Efficient and safe delivery of CRISPR components is crucial for therapeutic success.
Calcium and chitosan possess advantageous physicochemical properties for biomaterial applications.

Purpose of the Study:

To investigate and optimize the use of calcium-chitosan composites for plasmid CRISPR (pCRISPR) delivery.
To explore the efficacy of various calcium forms (nanoparticles, phosphate) combined with chitosan.
To evaluate the safety and stability of these delivery systems using green synthesis methods.

Main Methods:

Preparation of calcium nanoparticles (CaNPs), calcium phosphate (CaP), and their blends with chitosan (CaNPs/Chitosan, CaP/Chitosan, CaNPs-CaP/Chitosan).
Utilized routine and green synthesis procedures, including Salvia hispanica extract.
Tested pCRISPR delivery in a human embryonic kidney (HEK-293) cell line.

Main Results:

Optimized calcium-chitosan formulations demonstrated acceptable DNA binding for pCRISPR delivery.
CaP/Chitosan blend enhanced green fluorescent protein (EGFP) expression by approximately 25%.
CaNPs-CaP/Chitosan blend showed significant EGFP enhancement (over 14%) and nanoparticle stability.

Conclusions:

Calcium-chitosan composites are effective and stable carriers for pCRISPR delivery.
Green synthesis methods enhance nanoparticle stability and reduce potential toxicity.
These findings support the development of novel biomaterials for gene therapy applications.