A genetic strategy to treat sickle cell anemia by coregulating globin transgene expression and RNA interference

Nat Biotechnol. 2006 Jan;24(1):89-94. doi: 10.1038/nbt1176. Epub 2005 Dec 25.


The application of RNA interference (RNAi) to stem cell-based therapies will require highly specific and lineage-restricted gene silencing. Here we show the feasibility and therapeutic potential of coregulating transgene expression and RNAi in hematopoietic stem cells. We encoded promoterless small-hairpin RNA (shRNA) within the intron of a recombinant gamma-globin gene. Expression of both gamma-globin and the lariat-embedded small interfering RNA (siRNA) was induced upon erythroid differentiation, specifically downregulating the targeted gene in tissue- and differentiation stage-specific fashion. The position of the shRNA within the intron was critical to concurrently achieve high-level transgene expression, effective siRNA generation and minimal interferon induction. Lentiviral transduction of CD34(+) cells from patients with sickle cell anemia led to erythroid-specific expression of the gamma-globin transgene and concomitant reduction of endogenous beta(S) transcripts, thus providing proof of principle for therapeutic strategies that require synergistic gene addition and gene silencing in stem cell progeny.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anemia, Sickle Cell / genetics*
  • Anemia, Sickle Cell / metabolism*
  • Anemia, Sickle Cell / pathology
  • Anemia, Sickle Cell / therapy
  • Cells, Cultured
  • Gene Expression Regulation / genetics
  • Gene Silencing
  • Gene Targeting / methods
  • Genetic Therapy / methods*
  • Globins / genetics*
  • Globins / metabolism*
  • Globins / therapeutic use
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • RNA Interference*
  • Treatment Outcome


  • Globins