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CRISPR Gene Therapy

Investigating gene editing approaches to correct G6PD mutations and potentially cure G6PD deficiency at its genetic source.

Interactive Gene Editor

Simulating correction of the G6PD Mediterranean (S188F) mutation

Wild Type Sequence (Codon 188)
Mutant Sequence (S188F)
Amino Acid
Wild: Serine (S) Mutant: Phenylalanine (F)
Status: Ready

How CRISPR-Cas9 Works

The molecular scissors that enable precise genome editing

Cas9
Guide RNA

Targets specific DNA sequence

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Gene Editing Approaches

Multiple strategies for correcting G6PD mutations

CRISPR-Cas9

Creates double-strand breaks for gene knockout or HDR-mediated correction.

Pros
  • • Well-established
  • • High efficiency
  • • Versatile applications
Cons
  • • Off-target effects
  • • DSB-related risks
  • • Delivery challenges

Base Editing

Directly converts one base to another without double-strand breaks.

Pros
  • • No DSBs
  • • Precise single-base
  • • Lower off-targets
Cons
  • • Limited to C→T, A→G
  • • Bystander editing
  • • Size constraints

Prime Editing

"Search and replace" editing that can make any substitution or small indels.

Pros
  • • All 12 transitions
  • • No DSBs or donors
  • • Highly precise
Cons
  • • Lower efficiency
  • • Large cargo size
  • • Still emerging

Research Roadmap

Path from laboratory research to potential clinical application

Phase 1: Target Design

Design guide RNAs targeting common G6PD mutations. Computational prediction of off-target sites using Cas-OFFinder and CRISPOR.

Phase 2: In Vitro Testing

Test editing efficiency in patient-derived cells. Validate correction of enzyme activity using G6PD activity assays.

Phase 3: Delivery Optimization

Develop LNP or viral delivery systems for hematopoietic stem cells. Optimize for bone marrow targeting.

Phase 4: Preclinical Studies

Animal model studies to assess safety, biodistribution, and long-term efficacy of gene correction.

Key Challenges

  • Delivery: Efficiently reaching hematopoietic stem cells
  • Specificity: Avoiding off-target genome modifications
  • Durability: Ensuring long-term correction in self-renewing cells
  • Safety: Minimizing immunogenicity and genotoxicity
  • Scalability: Manufacturing for clinical application

Ethical Considerations

Gene therapy for G6PD raises important bioethical questions about germline editing, access equity, and informed consent. My research includes studying these ethical frameworks alongside the science.

My Current Work

Computational Guide RNA Design

I'm currently using computational tools to design and evaluate guide RNAs for correcting the G6PD Mediterranean mutation using base editing.

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Guides Designed
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High-Specificity
3
Top Candidates