Cas9, Cpf1 and C2c1/2/3-What's next?

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• The clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system, emerging in 2012-2013, revolutionized genome engineering.
• CRISPR-Cas9 is an RNA-guided DNA endonuclease utilizing Cas9 nuclease and a customizable single guide RNA for precise genomic targeting.

Purpose of the Study:

• To review advancements and alternatives to the standard CRISPR-Cas9 system.
• To highlight improvements addressing limitations of the original Streptococcus pyogenes Cas9 (SpCas9).

Main Methods:

• Discussion of engineered Cas9 variants with altered properties.
• Exploration of Cas9 homologs and novel Cas proteins beyond Cas9.
• Review of applications including gene knockdown and RNA-targeting imaging.

Main Results:

• Engineered Cas9 variants and homologs offer enhanced efficiency and specificity.
• Novel Cas proteins expand the toolkit for genome engineering.
• CRISPR-Cas systems provide flexible control for multiple gene loci and transcript imaging.

Conclusions:

• CRISPR-Cas technology continues to evolve with diverse systems offering improved genome engineering capabilities.
• These advancements facilitate precise genetic manipulation, gene knockdown, and advanced imaging techniques.