1. Volume 15, Issue 4, Pages 801-809 (April 2007)
Suppression of Clonal Dominance in Cultured Human Lymphoid Cells by Addition of the cHS4 Insulator to a Lentiviral Vector 
Marguerite V Evans-Galea, Matthew M Wielgosz, Hideki Hanawa, Deo Kumar Srivastava, Arthur W Nienhuis 
Molecular Therapy 
Volume 15, Issue 4, Pages (April 2007)
DOI: /sj.mt
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

2. Figure 1
Experimental strategy and vector design. (a) Derivation of single cell isolates of GFP-expressing Jurkat T cells. Transductions were performed at relatively low multiplicities of infection to enhance the probability that the cell populations derived from individual cell isolates would contain only a single copy of a vector genome. (b) Diagram of the two vectors used in these studies. The vector cwINS included a 1.2-kb fragment containing hypersensitive site 4 (HS4) from 5' of the chicken β-globin gene cluster. Shown is the inte-grated proviral genome in which both LTRs contain the insulator element. The solid bars indicate the location of probes used in Southern blot analysis. cPPT, central polypurine tract; G/P, residual retroviral sequences encoding GAG or POL; GFP, coding sequence for green fluorescent protein; Mp, DNA fragment containing the enhancer/promoter from the murine stem cell virus oncoretrovirus; RRE, rev-responsive element; WPRE, woodchuck post-transcriptional regulatory element. The location of the primers used for integration site mapping with the LM-PCR methodology are shown.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

3. Figure 2
Southern blot analysis of individual Jurkat T-cell isolates. (a) Vector copy-number determination by restriction with an enzyme that cuts once in the proviral genome (Figure 1b). (b) Evaluation of vector integrity using an enzyme that cuts twice in the proviral genome (Figure 1b).
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

4. Figure 3
Distribution of integration sites of the two vectors on the human chromosomes. The control cw unique integration sites are shown by the open circles on the right-hand side of each chromosome, and the unique integration sites of the vector containing the insulator, cwINS, are shown by the shaded circles on the left-hand side of each chromosome.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

5. Figure 4
Distribution of unique integration sites. (a) Distribution of sites with the two vectors with respect to the nearest transcriptional start site (TSS). (b) Distribution of the unique integration sites within transcriptional units for the two vectors.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions

6. Figure 5
Comparison of the number of sets of isolates with the same unique integration site for the two vectors. For example, one set of 12 and one set of 10 isolates with the same integration site were defined for the cw vector.
Molecular Therapy  , DOI: ( /sj.mt )
Copyright © 2007 The American Society of Gene Therapy Terms and Conditions