Insert, remove or replace: A highly advanced genome editing system using CRISPR/Cas9.

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) system, initially identified in prokaryotes, is now a leading genome editing tool.
• Its popularity stems from the Cas9 protein's ability to create targeted DNA double-strand breaks, guided by short RNA molecules, offering a simpler and more economical alternative to TALENs and ZFNs.

Purpose of the Study:

• To review recent advancements in the CRISPR/Cas9 system for genome editing in diverse species.
• To highlight available software tools for designing CRISPR/Cas9 targeting plasmids.
• To discuss the future potential of CRISPR/Cas9 technology in life science research, including cancer research.

Main Methods:

• Literature review of CRISPR/Cas9 applications in prokaryotes, fungi, plants, and animals.
• Analysis of the mechanism enabling precise DNA alterations.
• Identification and discussion of computational tools for experimental design.

Main Results:

• The CRISPR/Cas9 system has demonstrated increasing sophistication, ease of use, and applicability across a wide range of species.
• Significant progress has been made in applying CRISPR/Cas9 for genome editing in prokaryotes, fungi, plants, and animals.
• Various software tools are now available to facilitate the design of CRISPR/Cas9 targeting plasmids.

Conclusions:

• The CRISPR/Cas9 system is a powerful and versatile technology poised to address complex challenges in molecular biology and life science research.
• Continued advancements are expected to expand its utility and impact, particularly in areas like cancer research.
• The accessibility and efficiency of CRISPR/Cas9 facilitate its widespread adoption and innovation in genetic engineering.