Adenine Base Editor C-terminal Fragment

C-terminal fragment of ABE7.10 split via Npu intein at C574. Pairs with N-terminal fragment for editor reconstitution.

Length: 2385 bp (795 aa)

Type: Adenine Base Editor

Conversion: A-to-G

Editing window: Positions 4-7

Origin: SpCas9 D10A nickase (amino acids 574-1368) with Npu intein C-terminal domain

Characteristics

Contains C-terminal portion of SpCas9 D10A (residues 574-1368) including PAM recognition and DNA binding domains. Split site at C574 enables Npu intein trans-splicing with N-terminal fragment. Reference AAV construct includes U6-sgRNA expression cassette with BsmBI cloning sites for protospacer insertion and RFP dropout selection. Cbh promoter drives Cas9 fragment expression.

Applications: Dual-delivery base editing systems requiring split architecture. Includes sgRNA expression for single-vector guide RNA delivery in C-terminal construct. Validated for AAV-mediated delivery across multiple tissues. Can be adapted to other delivery platforms when combined with N-terminal fragment.

Limitations: Non-functional without N-terminal pair. Total coding sequence of reconstituted editor approaches AAV packaging limits. Requires BsmBI digestion and ligation to insert target-specific protospacer sequences in reference construct. Must be co-delivered with split-abe-nt for A-to-G editing activity.

Architecture: C-terminal SpCas9 D10A (residues 574-1368) with Npu intein C-terminal fragment at C574

Sequence

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References

Gaudelli NM, Komor AC, Rees HA, et al. (2017). Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464-471 - Gaudelli 2017 Adenine Base Editor
Levy et al. (2020). Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses. Nat Biomed Eng - Levy 2020 BE3.9max