CRISPR is revolutionizing the field of gene editing. It has the potential to alter the course of research and drug discovery, by providing scientists with a powerful tool to change any gene, in any cell in a highly targeted manner and without introducing foreign DNA. The benefits of CRISPR/Cas9 over previous forms of gene editing, such as TALENs and zinc finger nuclease (ZFN), are that it is much simpler to implement and has higher efficiency at performing bi-allelic gene modifications.

CRISPR/Cas9 genome editing has become widely used due to its simplicity and versatility, and the CRISPR technology has been adapted for diverse applications aside from genome editing. As a leader in gene synthesis and genome editing, CELLFREE offers validated CRISPR products, services and resources to help you harness the power of CRISPR genome editing for your research.

Introduction

CRISPR gene editing is a method by which the genomes of living organisms may be edited. It is based on a simplified version of the bacterial CRISPR/Cas (CRISPR-Cas9) antiviral defense system. By delivering the Cas9 nuclease complexed with a synthetic guide RNA (gRNA) into a cell, the cell's genome can be cut at a desired location, allowing existing genes to be removed and/or new ones added. The Cas9-gRNA complex corresponds with the CAS III CRISPR-RNA complex in the accompanying diagram.

While genomic editing in eukaryotic cells has been possible using various methods since the 1980s, the methods employed had proved to be inefficient and impractical to implement on a larger scale. Genomic editing leads to irreversible changes to the gene. Working like genetic scissors, the Cas9 nuclease opens both strands of the targeted sequence of DNA to introduce the modification by one of two methods. Knock-in mutations, facilitated via Homology Directed Repair (HDR), is the traditional pathway of targeted genomic editing approaches. This allows for the introduction of targeted DNA damage and repair. HDR employs the use of similar DNA sequences to drive the repair of the break via the incorporation of exogenous DNA to function as the repair template. This method relies on the periodic and isolated occurrence of DNA damage at the target site in order for a repair to commence. Knock-out mutations caused by Cas9/CRISPR results in the repair of the double-strand break by means of NHEJ (Non-Homologous End Joining). NHEJ can often result in random deletions or insertions at the repair site disrupting or altering gene functionality. Therefore, genomic engineering by CRISPR-Cas9 allows researchers the ability to generate targeted random gene disruption.

Competitive Advantages of CRISPR-Cas9

• Applicable in any type of cell lines
• Simultaneously knockout multiple targets
• More efficient
• Fast turnaround time

Delivery Form

Certificate of analysis (COA) for project

Lyophilized plasmid DNA (2~5 µg), or libraries

Sequencing chromatogram

QC report

*Pricing does not include the additional cost for gene synthesis, if required.

CRISPR Workflow