ZFIN ID: ZDB-PUB-200719-2
BE4max and AncBE4max Are Efficient in Germline Conversion of C:G to T:A Base Pairs in Zebrafish
Carrington, B., Weinstein, R.N., Sood, R.
Date: 2020
Source: Cells   9(7): (Journal)
Registered Authors: Sood, Raman
Keywords: AncBE4max, BE4max, base editing, genome editing, zebrafish
MeSH Terms:
  • Animals
  • Base Pairing/genetics*
  • Base Sequence
  • Cytosine/metabolism
  • Gene Editing*
  • Genetic Testing
  • Germ Cells/metabolism*
  • Zebrafish/genetics*
PubMed: 32674364 Full text @ Cells
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ABSTRACT
The ease of use and robustness of genome editing by CRISPR/Cas9 has led to successful use of gene knockout zebrafish for disease modeling. However, it still remains a challenge to precisely edit the zebrafish genome to create single-nucleotide substitutions, which account for ~60% of human disease-causing mutations. Recently developed base editing nucleases provide an excellent alternate to CRISPR/Cas9-mediated homology dependent repair for generation of zebrafish with point mutations. A new set of cytosine base editors, termed BE4max and AncBE4max, demonstrated improved base editing efficiency in mammalian cells but have not been evaluated in zebrafish. Therefore, we undertook this study to evaluate their efficiency in converting C:G to T:A base pairs in zebrafish by somatic and germline analysis using highly active sgRNAs to twist and ntl genes. Our data demonstrated that these improved BE4max set of plasmids provide desired base substitutions at similar efficiency and without any indels compared to the previously reported BE3 and Target-AID plasmids in zebrafish. Our data also showed that AncBE4max produces fewer incorrect and bystander edits, suggesting that it can be further improved by codon optimization of its components for use in zebrafish.
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