VCN ENHANCER COMPOSITIONS AND METHODS OF USING THE SAME

1 .- 326 . (canceled)

327 . A method of transducing a population of CD34 + hematopoietic stem or progenitor cells comprising culturing the cells in a culture medium, in the presence of a lentiviral vector, prostaglandin E 2 (PGE 2 ) or 16,16-dimethyl PGE 2 , and poloxamer 338, wherein the population of cells is transduced at least about 2 hours.

328 . The method of claim 327 , wherein the hematopoietic stem or progenitor cells are selected for CD34 + expression prior to transduction.

329 . The method of claim 327 , wherein the CD34 + hematopoietic stem or progenitor cells comprise the β-globin alleles selected from the group consisting of: β E /β 0 , β C /β 0 , β 0 /β 0 , β E /β E , β C /β + , β E /β + , β 0 /β + , β + /β + , β C /β C , β E /β S , β 0 /β S , β C /β S , β + /β S and β S /β S .

330 . The method of claim 327 , wherein the CD34+ hematopoietic stem or progenitor cells comprise the β-globin alleles selected from the group consisting of: β E /β 0 , β 0 /β 0 , β E /β E , β E /β + , β 0 /β + , and β + /β + .

331 . The method of claim 327 , wherein the lentiviral vector is cultured with the cells at an MOI of about 10 to about 30.

332 . The method of claim 327 , wherein the lentiviral vector is cultured with the cells at an MOI of about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29 or about 30.

333 . The method of claim 327 , wherein the lentiviral vector is derived from a lentivirus selected from the group consisting of: HIV (human immunodeficiency virus; including HIV type 1, and HIV type 2); visna-maedi virus (VMV) virus; the caprine arthritis-encephalitis virus (CAEV); equine infectious anemia virus (EIAV); feline immunodeficiency virus (FIV); bovine immune deficiency virus (BIV); and simian immunodeficiency virus (SIV)

334 . The method of claim 327 , wherein the lentiviral vector is derived from an HIV-1 lentivirus.

335 . The method of claim 327 , wherein the lentiviral vector comprises:

a) a 5′ long terminal (LTR);

b) a Psi (Ψ) packaging signal;

c) an RNA export element;

d) a lentiviral central polypurine tract (cPPT);

e) a promoter operably linked to a polynucleotide of interest; and

f) a SIN 3′ LTR.

336 . The method of claim 327 , wherein the lentiviral vector comprises:

a) a 5′ long terminal (LTR) comprising a cytomegalovirus (CMV) promoter;

b) a Psi (Ψ) packaging signal;

c) a Rev response element (RRE);

d) a lentiviral central polypurine tract (cPPT);

e) a human β-globin LCR and human β-globin promoter operably linked to a polynucleotide encoding a globin; and

f) a self-inactivating (SIN) 3′ LTR that comprises a synthetic polyadenylation sequence.

337 . The method of claim 336 , wherein the globin is selected from the group consisting of: a human β-globin, a human δ-globin, a human γ-globin, a human β A-T87Q -globin, a human β A-G16D/E22A/T87Q -globin, and a human β A-T87Q/K95E/K120E -globin.

338 . The method of claim 327 , wherein the lentiviral vector is an AnkT9W vector, a T9Ank2W vector, a TNS9 vector, a lentiglobin HPV569 vector, a lentiglobin BB305 vector, a BG-1 vector, a BGM-1 vector, a d432βAγ vector, a mLARβΔγV5 vector, a GLOBE vector, a G-GLOBE vector, a βAS3-FB vector, a V5 vector, a V5m3 vector, a V5m3-400 vector, a G9 vector, or a BCL11A shmir vector.

339 . The method of claim 327 , wherein the cells are cultured in the presence of PGE 2 .

340 . The method of claim 327 , wherein the cells are cultured in the presence of 16,16-dimethyl PGE 2 .

341 . The method of claim 327 , wherein the PGE 2 or 16,16-dimethyl PGE 2 is present in the culture in a concentration of about 10 μM to about 100 μM.

342 . The method of claim 327 , wherein the PGE 2 or 16,16-dimethyl PGE 2 is present in the culture in a concentration of about 10 μM to about 50 μM.

343 . The method of claim 327 , wherein the poloxamer 338 is present in the culture in a concentration of about 50 μg/mL to about 1000 μg/mL.

344 . The method of claim 327 , wherein the poloxamer 338 is present in the culture in a concentration of about 62.5 μg/mL to less than 1000 μg/mL.