Recombinant lentiviral vector for stem cell-based gene therapy of sickle cell disorder
AssigneesInstitut National De La Sante Et De La Recherche Medicale (Inserm), Universite Paris Descartes, Assistance Publique-Hopitaux de Paris, Imagine Institut Des Maladies Genetiques Necker Enfants Malades
InventorsAnnarita MICCIO, Vasco MENEGHINI
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Abstract
This invention relates to recombinant lentiviral vectors, compositions thereof, the use of the vectors or the compositions thereof, kits of parts comprising said vectors or compositions thereof and a catalytically active Cas9 or Cpf1 protein, methods for modifying the genome of a hematopoietic stem/progenitor cell (HSPC), and the HSPC obtainable by such methods.
Claims8
A genetically modified hematopoietic stem/progenitor cell (HSPC) obtainable by a method for modifying the genome of a HSPC, in vitro or ex vivo, comprising the steps of:
- a) contacting a HSPC with a recombinant lentiviral vector to obtain a transduced HSPC, wherein the lentiviral vector is integrated into the genome of said HSPC;
- and
- b) introducing into the transduced HSPC a catalytically active Cas9 or Cpf1 protein or a nucleotide sequence encoding a catalytically active Cas9 or Cpf1 protein, said catalytically active Cas9 or Cpf1 protein disrupts the expression and/or the function of a target gene when introduced or expressed into the transduced HSPC,
- wherein the recombinant lentiviral vector comprises in its genome:
- (i) a nucleotide sequence encoding a protein that has a therapeutic effect, said protein being selected from the group consisting of beta-globin, gamma-globin, delta-globin and variants thereof;
- and
- (ii) a nucleotide sequence encoding a guide RNA (gRNA) that comprises a spacer adapted to bind to a target nucleotide sequence, said target nucleotide sequence is:
- a. within the coding sequence or within a transcribed non-coding sequence of a target gene, said target gene is selected from beta-globin gene and BCL11A gene, or
- b. within the promoter region of a target gene, said target gene is gamma-globin gene.
A genetically modified hematopoietic stem/progenitor cell (HSPC) obtainable by a method for preparing the genetically modified HSPC, in vitro or ex vivo, comprising the steps of:
- a) contacting a HSPC with a recombinant lentiviral vector to obtain a transduced HSPC, wherein the lentiviral vector is integrated into the genome of said HSPC;
- and
- b) introducing into the transduced HSPC a catalytically active Cas9 or Cpf1 protein or a nucleotide sequence encoding a catalytically active Cas9 or Cpf1 protein, said catalytically active Cas9 or Cpf1 protein disrupts the expression and/or the function of a target gene when introduced or expressed into the transduced HSPC,
- wherein the recombinant lentiviral vector comprises in its genome:
- (i) a nucleotide sequence encoding a protein that has a therapeutic effect, said protein being selected from the group consisting of beta-globin, gamma-globin, delta-globin and variants thereof;
- and
- (ii) a nucleotide sequence encoding a guide RNA (gRNA) that comprises a spacer adapted to bind to a target nucleotide sequence, said target nucleotide sequence is:
- a. within the coding sequence or within a transcribed non-coding sequence of a target gene, said target gene is selected from beta-globin gene and BCL11A gene, or
- b. within the promoter region of a target gene, said target gene is gamma-globin gene.
Dependencies are inferred from the claim text.