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. 2022 Oct 18;12:16952. doi: 10.1038/s41598-022-20970-6

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Knockout of TRIM5 sensitized MDBK cells to HIV-1 infection. (a) Genomic DNA of MDBK cells encoding TRIM5 was amplified using PCR. The amplicon was mixed with sgRNA and Cas9 protein to determine the cleavage efficiency. The results of electrophoresis are presented. “M” denotes marker, and “(-)” indicates an amplicon mixed with only Cas9 protein. (b) MDBK TRIM5-knockout (k/o) cells (clones #4–6) or normal MDBK cells (Control) were infected with HIV-1 vectors encoding luciferase reporter protein. The infectivity was determined as relative light units (RLU) 2 days after infection. Relative infectivity was calculated using the control values. The results are presented as the mean and standard deviation of triplicate measurements from one assay, and they are representative of at least three independent experiments. Differences were examined by a two-tailed, unpaired Student’s t-test. ***p < 0.001. (c) Genomic DNA of MDBK TRIM5 k/o cells (clones #4–6) was amplified using PCR. The amplicon was subjected to sequencing to identify gene editing. The raw data on 4Peaks software (Nucleobytes) are presented.