JoVE Journal

Immunology and Infection

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Assessing Somatic Hypermutation in Ramos B Cells after Overexpression or Knockdown of Specific Genes

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08:12 min

November 1st, 2011

November 1st, 2011

18,035 Views

0:05

Title

1:00

Transfecting BOSC 23 Cells and Harvesting Viral Particles

2:27

Infecting B Cells

3:34

Single Cell Seeding

4:42

Analysis: IgM Loss (3 Weeks and 5 Weeks)

5:57

Representative Effects of Overexpressing Activated-induced Cytidine Deaminase (AID) or Knocking Down a Repair Factor in Ramos Cells

7:48

Conclusion

We describe how to perform retroviral or lentiviral infections of overexpression or shRNA-containing constructs in the human Ramos B-cell line and how to measure somatic hypermutation in these cells.

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