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Cited by 320CRISPR and Genetic EngineeringRNA regulation and diseaseBacterial Genetics and Biotechnology

A Review on the Mechanism and Applications of CRISPR/Cas9/Cas12/Cas13/Cas14 Proteins Utilized for Genome Engineering

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Hillary, V. Edwin, Ceasar, S. Antony·2023·Molecular Biotechnology
DOI 10.1007/s12033-022-00567-0PMC PMC9512960
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Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (CRISPR/Cas) system has altered life science research offering enormous options in manipulating, detecting, imaging, and annotating specific DNA or RNA sequences of diverse organisms. This system incorporates fragments of foreign DNA (spacers) into CRISPR cassettes, which are further transcribed into the CRISPR arrays and then processed to make guide RNA (gRNA). The CRISPR arrays are genes that encode Cas proteins. Cas proteins provide the enzymatic machinery required for acquiring new spacers targeting invading elements. Due to programmable sequence specificity, numerous Cas proteins such as Cas9, Cas12, Cas13, and Cas14 have been exploited to develop new tools for genome engineering. Cas variants stimulated genetic research and propelled the CRISPR/Cas tool for manipulating and editing nucleic acid sequences of living cells of diverse organisms. This review aims to provide detail on two classes (class 1 and 2) of the CRISPR/Cas system, and the mechanisms of all Cas proteins, including Cas12, Cas13, and Cas14 discovered so far. In addition, we also discuss the pros and cons and recent applications of various Cas proteins in diverse fields, including those used to detect viruses like severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). This review enables the researcher to gain knowledge on various Cas proteins and their applications, which have the potential to be used in next-generation precise genome engineering.

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  • Several Cas variants, such as Cas12 and Cas13, have been developed to enhance DNA cleavage and expand genome editing versatility.

    “In recent years, several Cas variants, such as Cas12 and Cas13, have been developed to enhance DNA cleavage and expand genome editing versatility [7,8].”

    From Precise CRISPR/Cas9 and Cas12 Correction Using Lipoplexes in Retinal Models Derived from Patients with Inherited Retinal Dystrophies (2026) · cites this paper as [8]

    The Introduction says Cas12 improves genome-editing efficiency and that Cas12 and Cas13 broaden editing applications. The Cas12 Protein > Mechanism section describes DNA cleavage, but the Cas13 Protein > Mechanism of Cas13a Protein section describes cleavage of RNA, not DNA. Thus, the paper does not support the implication that both variants enhance DNA cleavage. (LLM, conf=0.88)

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Cited by

Sorted by citations

Authors
Hillary, V. E., & Ceasar, S. A.
Affiliations
Rajagiri College of Social Sciences (Autonomous) (IN)
Corresponding author
Ceasar, S. Antony
ORCID
Venue
Molecular Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2022-09-27
Volume
65
Issue
3
Pages
311-325
ISSN
  • 1073-6085 (ISSN-L)
  • 1559-0305
DOI
10.1007/s12033-022-00567-0
PMC
PMC9512960
OpenAlex
W4297102731
Cited by
320
References
5
Citations by year
2026: 74 · 2025: 126 · 2024: 88 · 2023: 32
FWCI
6.40
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Biotechnology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
CRISPR-Cas, Cas9, Cas12, Cas13, genome editing, guide RNA, CRISPR spacers, COVID-19 detection, RNA editing
MeSH terms
Gene Editing, COVID-19, SARS-CoV-2, DNA, Humans, CRISPR-Cas Systems

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