Abstract: In certain embodiments an optimized derivative of the CCLc-?AS3-FB lentiviral vector termed (CCLc-mGata/ANK-CoreLCR-?AS3-FB), is provided which is capable of driving lineage-restricted expression of a beta-globin gene (e.g., an anti-sickling ?-globin like gene (?A83)). In certain embodiments the vectors described herein comprise novel defined LCR HS core sequences (HS2(˜420 bp), HS3?40 bp), HS4(˜410 bp)) which can be used to replace the putative LCR HS sequences present within the “mini-LCR” (˜3.6 kb reduced to ˜1.2 kb) to produce an “optimized mini-LCR”.

Abstract: In certain embodiments methods of treating X-Linked agammaglobulinemia (XLA) in a mammal are provided where the methods comprise: i) providing differentiated T cells and/or stem/progenitor cells from the mammal; ii) performing a targeted insertion of a corrective BTK cDNA at the BTK gene locus in said cells to provide a corrected BTK gene in said cells; and iii) introducing said cells into said mammal where said corrected BTK gene is expressed in a physiologically regulated manner.

Abstract: In certain embodiments an optimized derivative of the CCLc-?AS3-FB lentiviral vector termed (CCLc-mGata/ANK-CoreLCR-?AS3-FB), is provided which is capable of driving lineage-restricted expression of a beta-globin gene (e.g., an anti-sickling ?-globin like gene (?A83)). In certain embodiments the vectors described herein comprise novel defined LCR HS core sequences (HS2(˜420 bp), HS3?40 bp), HS4(˜410 bp)) which can be used to replace the putative LCR HS sequences present within the “mini-LCR” (˜3.6 kb reduced to ˜1.2 kb) to produce an “optimized mini-LCR”.

Abstract: In various embodiments a recombinant lentiviral vector is provided comprising an expression cassette comprising a nucleic acid construct comprising an anti-sickling human beta globin gene encoding an anti-sickling-beta globin polypeptide comprising the mutations Gly16Asp, Glu22Ala and Thr87Gln, where the lentiviral vector is a TAT-independent and self-inactivating (SIN). In certain embodiments the vector additionally contains one or more insulator elements. The vectors are useful in gene therapy for the treatment of sickle cell disease.