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Review
. 2024 Dec 12:6:1509924.
doi: 10.3389/fgeed.2024.1509924. eCollection 2024.

Advances in CRISPR-Cas technology and its applications: revolutionising precision medicine

Sarkar Sardar Azeez  1 , Rahin Shareef Hamad  2 , Bahra Kakamin Hamad  3 , Mudhir Sabir Shekha  4   5 , Peter Bergsten  5
Affiliations
Review

Advances in CRISPR-Cas technology and its applications: revolutionising precision medicine

Sarkar Sardar Azeez et al. Front Genome Ed. .

Abstract

CRISPR-Cas (Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-associated proteins) has undergone marked advancements since its discovery as an adaptive immune system in bacteria and archaea, emerged as a potent gene-editing tool after the successful engineering of its synthetic guide RNA (sgRNA) toward the targeting of specific DNA sequences with high accuracy. Besides its DNA editing ability, further-developed Cas variants can also edit the epigenome, rendering the CRISPR-Cas system a versatile tool for genome and epigenome manipulation and a pioneering force in precision medicine. This review explores the latest advancements in CRISPR-Cas technology and its therapeutic and biomedical applications, highlighting its transformative impact on precision medicine. Moreover, the current status of CRISPR therapeutics in clinical trials is discussed. Finally, we address the persisting challenges and prospects of CRISPR-Cas technology.

Keywords: CRISPR-Cas9; and CRISPR in clinical trials; cancer immunotherapy; epigenome modulation; genome editing.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

None
Graphical abstract
FIGURE 1
FIGURE 1
Schematic representation of CRISPR-Cas systems, categorised into Class 1 and Class 2. Each class is further divided into types and subtypes, showing the associated complexes and their primary targets.
FIGURE 2
FIGURE 2
This figure displays a range of Cas protein variants and engineered tools derived from the CRISPR-Cas family, highlighting their diverse applications in genome and transcriptome editing. Central to the diagram is the native Cas9-sgRNA complex, which serves as a foundation for multiple modifications. Shown clockwise from the top left, base and prime editors (e.g., cytosine and adenine base editors, prime editors) allow single-nucleotide changes and precise insertions without DSBs. Recombinant Cas9 proteins (Cas9-Rad51/Rad52 fusions) enhance homology-directed repair (HDR) outcomes, while high-fidelity Cas9 variants reduce off-target effects to improve targeting specificity. Transcriptional activators (CRISPRa) and repressors (CRISPRi) enable gene expression modulation without altering DNA sequences, using dCas9 fused to transcriptional regulators. The dCAS9-APEX2 complex, tags nearby proteins with biotin, which is useful in proteome mapping. PAM-expanded Cas9 variants (e.g., SpRY, SpCas9-NG) target a broader range of PAM sequences, increasing flexibility in target selection. In the lower-left corner, “Natural Cas9 Variants” encompass Cas12, which produces sticky-end DNA cuts; Cas13, an RNA editor targeting RNA without modifying DNA; and Cas14, a small-size protein that cuts single-stranded DNA, independent of PAM recognition. This diverse toolkit illustrates the flexibility and breadth of CRISPR-Cas systems for targeted genetic and epigenetic modifications across various applications.
FIGURE 3
FIGURE 3
Distribution of CRISPR-Cas clinical trials by therapeutic area.
See this image and copyright information in PMC

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