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Engineered prime editors with PAM flexibility
- Source :
- Molecular Therapy. 29:2001-2007
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Although prime editors are a powerful tool for genome editing, which can generate various types of mutations such as nucleotide substitutions, insertions, and deletions in the genome without double-strand breaks or donor DNA, the conventional prime editors are still limited to their target scopes because of the PAM preference of the Streptococcus pyogenes Cas9 (spCas9) protein. Here, we describe the engineered prime editors to expand the range of their target sites using various PAM-flexible Cas9 variants. Using the engineered prime editors, we could successfully generate more than 50 types of mutations with up to 51.7% prime-editing activity in HEK293T cells. In addition, we successfully introduced the BRAF V600E mutation, which could not be induced by conventional prime editors. These variants of prime editors will broaden the applicability of CRISPR-based prime editing technologies in biological research.
- Subjects :
- Proto-Oncogene Proteins B-raf
Computer science
Computational biology
medicine.disease_cause
Genome
Prime (order theory)
chemistry.chemical_compound
03 medical and health sciences
0302 clinical medicine
Genome editing
CRISPR-Associated Protein 9
Drug Discovery
Genetics
medicine
Humans
CRISPR
Clustered Regularly Interspaced Short Palindromic Repeats
Nucleotide Motifs
Molecular Biology
Alleles
030304 developmental biology
Gene Editing
Pharmacology
Flexibility (engineering)
0303 health sciences
Mutation
Binding Sites
Cas9
BRAF V600E
HEK293 Cells
chemistry
Amino Acid Substitution
030220 oncology & carcinogenesis
Molecular Medicine
CRISPR-Cas Systems
Genetic Engineering
DNA
Subjects
Details
- ISSN :
- 15250016
- Volume :
- 29
- Database :
- OpenAIRE
- Journal :
- Molecular Therapy
- Accession number :
- edsair.doi.dedup.....2d7a5226cd3a227b9692f17fcbb95fca