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. 2015 Feb 27;5(1):e1023423. doi: 10.1080/21592799.2015.1023423

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

© © Han B Lee, Zachary L Sebo, Ying Peng, and Yi Guo

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 2. — w2 TALEN-introduced somatic mutations and germline transmission. (A) The scheme of eye-color change introduced by w2 TALEN. From parental red-eyed flies (P), embryos injected w2 TALEN developed to G0 adults with mosaic eyes containing white patches. In G1 generation, white-eyed flies carried germline-transmitted mutations. (B) G0 somatic mutation rates at various w2 TALEN concentrations quantified by mosaic-eye phenotype. Numbers of G0 flies counted are listed on top of each bar. (C) The rational of quantifying global somatic mutations in individual G0 flies using SURVEYOR nuclease assay. Re-annealed amplicons around the predicted cut site undergone SURVEYOR nuclease treatment. The insert shows a representative result of digested product separation using Fragment Analyzer. Peaks at lower molecular weights (green) represent shorter DNA fragments generated by nuclease cut at a specific mutated site. See Figure S1 for more analysis and quantifications of gene modification rate. (D) G1 germline transmission rates at various w2 TALEN concentrations. The rate was represented by the percentage of white-eyed flies among total number of G1 progeny at each condition. Numbers of G1 flies counted are listed on top of each bar. (E) Comparisons of G1 germline transmission rates between progenies of non-mosaic eye (black bars) and mosaic eye G0 flies (green bars) at various w2 TALEN concentrations. Numbers of G1 flies counted are listed on top of each bar. (F) G0 larval survival rates at various w2 TALEN concentrations. This rate was represented by the percentage of injected embryos developed into first-instar larvae within 48 hours post injection from either wild-type (black bars) or lig4¹⁶⁹ embryos (orange bars). Numbers of G0 embryos counted are listed on top of each bar.