Optimization of determinants for successful genetic correction of diseases, mediated by hematopoietic stem cells

Abstract

Methods and compositions disclosed herein generally relates to methods of determining minimum hematopoietic stem cell (HSC) chimerism and gene dosage for correction of a hematopoietic disease; in particular, in in vivo models. The invention also relates to modified lentiviral expression vectors for increase a viral titer and various methods for increasing such titers as well as expression vectors capable of enhancing such titers. The invention also relates to CHS4 chromatin insulator-derived functional insulator sequences. The invention further relates to methods for genetic correction of diseases or reducing symptoms thereof, such as sickle cell anemia, a lysosomal storage disease. The invention further relates to a method of improving and/or correcting one or more central nervous system (CNS) abnormalities caused by one or more lysosomal storage disease. The invention further relates to methods of improving titer in transfection-based bioreactor culture production or transfection-based production systems using eukaryotic cells.

1 . A modified self-inactivating (SIN) lentiviral vector for expressing a transgene of interest, the vector comprises:
 (a) a lentiviral vector backbone comprising lentiviral cis elements, which consists essentially of
 (i) a packaging signal (ψ), and 
 (ii) one or more of a rev response element (RRE), a 5′ portion of Gag, and an Env slice acceptor sequence, and 
   (b) a transgene of interest.   
     
     
         2 .- 26 . (canceled)