Insertion of Fragments of DNA: Gene Disruption and Replacement.

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Presentation transcript:

Insertion of Fragments of DNA: Gene Disruption and Replacement

Gene transfer in murine system Several different combinations of growth factors have been successfully used for supporting gene transfer into murine stem cells. Combination of interleukin 3 (IL-3), interleukin 6 (IL-6), and stem cell factor (SCF). Inclusion of recently discovered early acting growth factors such as t-3 ligand and megakaryocyte growth and development factor (MGDF)/thrombopoietin (TPO) have augmented the level of genetically modied cells. Treatment of animals with 5-uorouracil before marrow harvest to stimulate cycling of primitive cells, The co-culture of target cells directly on a layer of retroviral producer cells or other stromal support, The use of high titer (greater than 10 5 viral particles per ml) vector and Co-localization of vector and target cells using bronectin-coated dishes.

The T cell membrane (shown as the purple, semicircle double line) allows entry of HIV (in pink) into the cell through multiple cell receptors anchored on the membrane, including CCR5.

Use of Genetically Modified Stem Cells in Experimental Gene Therapies