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. 2019 Jul 9;8:e46767. doi: 10.7554/eLife.46767

Figure 8. KHNYN is necessary for CpG dinucleotides to inhibit HIV-1 genomic RNA expression.

Figure 8.

(A–C) HeLa Control CRISPR cells, ZAP-G1 CRISPR cells, KHNYN-G1 CRISPR bulk cells or two independent clones were transfected with 500 ng pHIV-1 or pHIV-1EnvCpG86-561 and 500 ng pGFP-FLAG. RNA was extracted from cell lysates (A–B) and media (C). Genomic RNA (gRNA) (A, C) or nef mRNA (B) abundance was quantified by qRT-PCR. The bar charts show the average value of three independent experiments normalized to the value obtained for HeLa Control CRISPR cells co-transfected with pHIV-1 and pGFP-FLAG. Data are represented as mean ± SD. *p<0.05 as determined by a two-tailed unpaired t-test. p-values for HIV-1EnvCpG86-561 genomic RNA in Control cell versus ZAP-G1, KHNYN-G1 Bulk, KHNYN-G1 Clone A, and KHNYN-G1 Clone B CRISPR cell lysates are 2.98 × 10−2, 3.70 × 10−2, 4.26 × 10−2, and 1.30 × 10−4, respectively. p-Values for HIV-1EnvCpG86-561nef mRNA in Control cell versus ZAP-G1, KHNYN-G1 Bulk, KHNYN-G1 Clone A, and KHNYN-G1 Clone B CRISPR cell lysates are 0.29, 0.54, 0.90 and 0.42, respectively. p-Values for HIV-1EnvCpG86-561 genomic RNA in Control cell versus ZAP-G1, KHNYN-G1 Bulk, KHNYN-G1 Clone A, and KHNYN-G1 Clone B CRISPR cell media are 1.65 × 10−2, 8.64 × 10−3, 2.82 × 10−3, and 3.54 × 10−4, respectively.