Fig. 3: Construction of AID2 cells using degron-tagged AID.
From: Monoclonal antibody generation by controlled immunoglobulin gene rearrangements
a Flowchart of the AID2 cell construction process. First, endogenous AID genes are homogeneously knocked out, then TIR1 expression vector is introduced followed by the transfection of degron-tagged AID expression vector. b Comparison of the gene conversion frequency of mp232 (left) and mp232-ΔAID (right). Sample sizes are n = 24 for mp232 and n = 23 for mp232-ΔAID. Long horizontal bars represent the mean, and their values are shown alongside. Error bars represent ± s.d. Differences were compared using a two-tailed t-test. c Schematic representation of x3 degron tagged AID. C-terminal tagged (“AID-degron”, top), N-terminal tagged (“degron-AID”, middle) and the version degron tag was inserted downstream of NLS (“NLS-degron-AID (ΔNLS)”, bottom). d, e Gene conversion activity of the C-terminal degron tagged GdAID (d) and mAID (e). Results for the representative four independent clones are shown. Population of mCherry positive cells are highlighted by red rectangles. Ratios of the mCherry positive cells (%) are shown alongside.
