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Single-Base Resolution: Increasing the Specificity of the CRISPR-Cas System in Gene Editing
- Source :
- Mol Ther
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- The CRISPR-Cas system holds great promise in the treatment of diseases caused by genetic variations. The Cas protein, an RNA-guided programmable nuclease, generates a double-strand break at precise genomic loci. However, the use of the clustered regularly interspersed short palindromic repeats (CRISPR)-Cas system to distinguish between single-nucleotide variations is challenging. The promiscuity of the guide RNA (gRNA) and its mismatch tolerance make allele-specific targeting an elusive goal. This review presents a meta-analysis of previous studies reporting position-dependent mismatch tolerance within the gRNA. We also examine the conservativity of the seed sequence, a region within the gRNA with stringent sequence dependency, and propose the existence of a subregion within the seed sequence with a higher degree of specificity. In addition, we summarize the reports on high-fidelity Cas nucleases with improved specificity and compare the standard gRNA design methodology to the single-nucleotide polymorphism (SNP)-derived protospacer adjacent motif (PAM) approach, an alternative method for allele-specific targeting. The combination of the two methods may be advantageous in designing CRISPR-based therapeutics and diagnostics for heterozygous patients.
- Subjects :
- Review
Computational biology
Polymorphism, Single Nucleotide
03 medical and health sciences
0302 clinical medicine
Genome editing
Drug Discovery
Genetic variation
Genetics
Humans
SNP
CRISPR
Guide RNA
Molecular Biology
030304 developmental biology
Gene Editing
Pharmacology
0303 health sciences
Nuclease
biology
Genome, Human
Palindrome
Genomics
Endonucleases
Protospacer adjacent motif
030220 oncology & carcinogenesis
biology.protein
Molecular Medicine
CRISPR-Cas Systems
Subjects
Details
- ISSN :
- 15250016
- Volume :
- 29
- Database :
- OpenAIRE
- Journal :
- Molecular Therapy
- Accession number :
- edsair.doi.dedup.....3acb93cfc1b48cf9c4dbe0d280f93b43