human ccr5  (ATCC)

Journal: Nature Communications

Article Title: CXCR4-tropic HIV-1 infection in an immunocompetent monkey model

doi: 10.1038/s41467-026-70209-5

Figure Lengend Snippet: cDNAs for the indicated genes were sequenced from 191 owl monkeys (diploid = 382 alleles determined for each gene). Synonymous polymorphisms were ignored, and alleles encoding unique protein “variants” are grouped by circles. Protein variant numbers were assigned in the order in which they were discovered. The size of each circle indicates the number of times alleles encoding each variant were observed, with reference to the size key given in each panel. Tick marks between circles indicate the number of amino-acid substitutions that differentiate each encoded protein variant from its neighbors. Graphics generated using PopART . A The results for receptor genes. Alleles encoding 18 unique CD4 variants were identified, and variant 1 was observed 101 times. Alleles encoding two unique CCR5 variants were identified, and variant 2 was observed only one time. We discovered only a single CXCR4 variant, although this gene was sequenced from only 15 individuals (diploid = 30 alleles) instead of 191. B The results for restriction factor genes. Alleles encoding 8 different Tetherin variants were identified, with variant 4 observed 104 times. 37 different TRIMCyp variants were identified, and no variant was found more than 27 times. 14 different APOBEC3G variants were identified, and variant 1 appeared 185 times. All alleles summarized in this figure have been recorded in GenBank, as described in the methods.

Article Snippet: Cf2Th cells (ATCC CRL-1430) were transduced to stably express human CCR5 alone (Genbank # NM_001394783 ), or with human (Genbank # NM_000616 ) or various owl monkey CD4 variants.

Techniques: Variant Assay, Generated

Journal: Nature Communications

Article Title: CXCR4-tropic HIV-1 infection in an immunocompetent monkey model

doi: 10.1038/s41467-026-70209-5

Figure Lengend Snippet: Cf2Th cells were transduced to stably express human CCR5 alone, or with human or various owl monkey CD4 variants ( X axis). Cf2Th cell lines were plated, exposed to an equivalent volume of HIV-1-EGFP, and 48 h later assayed for EGFP signal by flow cytometry. Values were normalized to the average signal on cells expressing human CD4. The limit of detection (dashed line) was set based on signal observed from uninfected cells. Data points represent six individual technical replicates, three each from two independent experiments. Solid bar indicates the mean. Source data are provided in the Source Data file.

Article Snippet: Cf2Th cells (ATCC CRL-1430) were transduced to stably express human CCR5 alone (Genbank # NM_001394783 ), or with human (Genbank # NM_000616 ) or various owl monkey CD4 variants.

Techniques: Stable Transfection, Flow Cytometry, Expressing

Journal: Nature Communications

Article Title: CXCR4-tropic HIV-1 infection in an immunocompetent monkey model

doi: 10.1038/s41467-026-70209-5

Figure Lengend Snippet: A Restriction of HIV-1 by owl monkey TRIMCyp variants. CRFK cells stably expressing 14 TRIMCyp variants were exposed to VSV G-pseudotyped HIV-1-GFP. The percentage of cells expressing GFP was normalized relative to cells not expressing any TRIMCyp. Experiments were performed in two biological replicates, each with two technical replicates; each point is an average of the 2 technical replicates from a biological replicate, and the line is the average. B Screen for mutations in HIV-1 that result in escape from owl monkey TRIMCyp. CRFK cells stably expressing one of two TRIMCyp variants (variant 30 is the most common in the colony) were transduced with an increasing volume of VSV G-pseudotyped HIV-1-GFP containing cyclophilin A-binding loop amino-acid substitutions (G89V , or V86A ) or loops from 25 diverse SIV/HIV lentiviruses as indicated at the top (full set of 25 shown in Figure. ). C Alignment of the cyclophilin A-binding loop of HIV-1 (GenBank # AF324493 ) and SIVrcm (GenBank # AF028608 ), which differ at four amino-acid positions. D HIV-1 replication on owl monkey cells. Hut78 (human) and OMK (owl monkey) cells, both expressing human CD4 and CCR5, were exposed to HIV-1 with or without the four substitutions re-constituting the cyclophilin A-binding loop from SIVrcm as in ( C ) at an MOI = 0.1. Growth curves were generated by measuring the concentration of HIV-1 capsid (CA) in supernatants at the indicated time points. For all panels, source data are provided in the Source Data file.

Article Snippet: Cf2Th cells (ATCC CRL-1430) were transduced to stably express human CCR5 alone (Genbank # NM_001394783 ), or with human (Genbank # NM_000616 ) or various owl monkey CD4 variants.

Techniques: Stable Transfection, Expressing, Variant Assay, Transduction, Binding Assay, Generated, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: The Chimeric Nuclease SpRYc Exhibits Highly Variable Performance Across Biological Systems

doi: 10.3390/ijms27010488

Figure Lengend Snippet: Editing efficiencies of SpCas9 and SpRYc across diverse PAM contexts at the CCR5 target in 293T-CD4-CCR5 clone 19. ( A ) Sequences of sgRNAs for SpCas9 and SpRYc targeting the third exon of human CCR5 gene. The 110-nucleotide sequence of Homo sapiens (Hs) CCR5 exon 3 is shown. The sgRNA «sgR5-2» (PAM CGG) is not displayed, as it is located 117 nucleotides upstream of the depicted region. ( B ) CCR5 KO efficiency for different PAMs measured by flow cytometry. Data are presented as mean ± standard deviation (SD). Statistical significance was evaluated using one-way ANOVA with Dunnett’s multiple comparison test: ns, not significant; ****, p < 0.0001.

Article Snippet: For immunofluorescence staining, cells were washed once with PBS and incubated with Alexa 647-labelled antibodies against CCR5 (#E-AB-F1418M, Elabscience, USA) diluted in PBS at 4 °C for 30 min. Then, cells were washed twice with PBS and analyzed using a CytoFLEX S flow cytometer (Beckman-Coulter, USA).

Techniques: Sequencing, Flow Cytometry, Standard Deviation, Comparison