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Aim 2. Are nicks efficient and safe initiating sites for targeted gene correction?

Aim 2. Are nicks efficient and safe initiating sites for targeted gene correction?. NHEJ G1 phase. homology-directed repair S phase. DSBs can cause translocations. Merge/DAPI. - H2AX. 25 20 15 10 5 0. 21-fold. Foci per nucleus. -. 0.4 21 1. I-AniI

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Aim 2. Are nicks efficient and safe initiating sites for targeted gene correction?

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  1. Aim 2. Are nicks efficient and safe initiating sites for targeted gene correction? NHEJ G1 phase homology-directed repair S phase DSBs can cause translocations.

  2. Merge/DAPI -H2AX 25 20 15 10 5 0 21-fold Foci per nucleus - 0.4 21 1 I-AniI Cleavase 80 60 40 20 0 I-AniI Nickase 0.1 13 0.3 -H2AX foci induced by cleavase but not nickase -H2AX: cleavase/nickase 46-fold Percent nuclear area __ nickase cleavase

  3. Cleavase causes "off-target" damage at reporters I-AniI I-AniI I-AniI PolyLacO H2-Kd CD4 eGFP H2-Kd+ CD4- eGFP trGFP I-AniI H2-Kd- CD4- 2-Kd CD4 Percent of cells H2-Kd-CD4+ CD4 Deletion Gene correction — Cleavase Nickase Correction Deletion GFP- I-AniI Donor template GFP+ PolyLacO

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