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Evolved riboflavin biosynthesis enzyme RibD as a novel nucleic acid deaminase for base editing.

Interests: Gene Therapy
Lead Inventor: Weixin Tang

SUMMARY

Enhanced precision in cytosine base editing by employing an evolved riboflavin biosynthesis enzyme RibD that converts targeted cytosines to thymines without intrinsic sequence-context specificity, thereby expanding genomic editing flexibility and enabling more adaptable gene modification tools

The Unmet Need: Limitations in current base editors due to sequence-context dependent editing specificity

  • Existing cytosine base editors (CBEs) typically derive from naturally occurring cytidine deaminases that possess high intrinsic sequence-context preferences, limiting editing efficiency and scope across diverse genetic sites and complicating precise, targeted genome modification. This specificity constraint reduces the effectiveness of base editors in addressing varied genetic disorders and hinders the development of broadly applicable therapeutic and research tools.

  • The rapid expansion of gene editing technologies and increasing demand for precise, versatile gene therapies have spurred efforts to develop novel base editors with improved substrate flexibility and controllable sequence specificity, creating an opportunity to engineer new classes of enzymes that circumvent these limitations.

The Proposed Solution: Evolved RibD enzyme repurposed as a nucleic acid cytosine deaminase with minimized sequence-context bias

  • The faculty inventor developed cytosine base editors derived from an evolved form of the Escherichia coli RibD enzyme, originally involved in riboflavin biosynthesis and naturally acting on small-molecule substrates, which has been directed-evolved to deaminate cytosines on single-stranded DNA.
  • This novel enzymatic platform differentiates from traditional CBEs by lacking inherent sequence-context specificity, potentially enabling more flexible and tunable recognition of DNA targets. 

ADVANTAGES

  • Expanded target sequence flexibility

  • Reduced intrinsic sequence-context bias

  • Novel enzyme class for base editing

  • Potential platform for engineered specificity

  • Compatibility with single-stranded DNA substrates

  • Foundation for next-generation gene editing tools

APPLICATIONS

  • Targeted correction of single-nucleotide genetic disorders (cystic fibrosis, sickle-cell anemia)

  • Development of context-specific base editors for precision therapeutics

  • Generation of mutant gene libraries for functional genomics studies