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. 2022 Nov 15;14(7):540–545. doi: 10.1093/procel/pwac052

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©The Author(s) 2022. Published by Oxford University Press on behalf of Higher Education Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 2. — hfCas12Max mediates high-efficiency gene editing ex vivo and in vivo. (A) Schematics of hfCas12Max gene editing in primary human cells. (B) Viability and indel activity of human CD3⁺ T cells following delivery of hfCas12Max RNPs with three different TRAC targeted crRNAs at 1.6 and 3.2 μmol/L, respectively (n = 2 or 3). NC represents blank control, untreated with RNP. (C) Representative flow cytometric analysis of edited CD3⁺ T cell 5 days after RNP delivery. NC represents blank control, untreated with RNP. (D) Schematics of in vivo non-liposome delivery containing IVT-mRNA, LNP packaging process. (E) Editing efficiency of LNP packaging with hfCas12Max mRNA and targeted Ttr crRNA at increased concentrations in N2a cells (n = 8). (F) Schematics of Ttr locus. (G) Indel rates of LNP packaging with hfCas12Max mRNA and targeted Ttr crRNA at three dose (0.1, 0.3, and 0.5 mpk) in C57 mouse (n = 6). (H) The A to G editing percentage of LNP packaging with dCas12i-ABE mRNA and targeted Ttr crRNA at 3 mpk in C57 mouse (n = 2).