Integrate CRISPR/Cas9 for protein expression of HLA-B*38:68Q via precise gene editing.

The determination of null- or low-expressed HLA alleles is clinically relevant in both hematopoietic stem cell transplantation and solid organ transplantation. We studied the expression level of a questionable (Q) HLA-B*38:68Q allele, which carries a 9-nucleotide (nt) deletion at codon 230-232 in exon 4 of HLA-B*38:01:01:01 using CRISPR/Cas9 gene editing technology. CRISPR/Cas9 gene editing of HLA-B*38:01:01:01 homozygous EBV B cell line resulted in one HLA-B*38:68Q/B*38:01:01:01 heterozygous and one HLA-B*38:68Q homozygous clone. Flow cytometric analysis of monoclonal anti-Bw4 antibody showed the protein expression of HLA-B*38:01:01:01 in homozygous cells was 2.2 fold higher than HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the expression of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 2.0 fold higher than HLA-B*38:68Q homozygous cells. The HLA-B*38:68Q expression was further confirmed using anti-B38 polyclonal antibody. Similarly, the expression of the HLA-B*38:01:01:01 homozygous cells was 1.5 fold higher than that of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 1.6 fold higher than that of HLA-B*38:68Q homozygous cells. The treatment of HLA-B*38:68Q homozygous cells with IFN-γ significantly increased its expression. In conclusion, we demonstrate that HLA-B*38:68Q is a low-expressing HLA allele. The CRISPR/Cas9 technology is a useful tool to induce precise gene editing in HLA genes to enable the characterization of HLA gene variants on expression and function

Capillary and viscous forces during CO2 flooding in tight reservoirs

In this study, the multiphase multicomponent Shan-Chen lattice Boltzmann method is employed to analyze the impact of capillary force on oil-CO2-water  fluid flow and enhanced oil recovery. Various sizes of the single throat are designed to  simulate the interaction between displacing and displaced phases as well as their mechanical  equilibrium. Several sensitivities are taken into account, such as wettability, miscibility,  interfacial tension, and pore aperture. Based on the objective reservoir conditions,  supercritical CO2 as an injection fluid is adopted to study the influence of different displacement  patterns on the mechanical equilibrium in both homogenous and heterogeneous porous media, in  which enhanced oil recovery is also quantitatively estimated. The results show that the  water-alternating-gas injection pattern reduces the moving speed of the leading edge by  increasing the swept area of the residual oil, and inhibits the breakthrough effect of the  gas, making it the optimal displacement method in terms of the degree of oil production. Compared with the results of different displacement patterns, the enhanced oil recovery  of water-alternatinggas injection is the highest, followed by supercritical CO2 flooding after  water flooding, and lastly, continuous supercritical CO2 flooding.Cited as: Zhang, C., Zhang, Q., Wang, W., Xie, Q., Su, Y., Zafar, A. Capillary and viscous forces during CO2 flooding in tight reservoirs. Capillarity, 2022, 5(6): 105-114. https://doi.org/10.46690/capi.2022.06.0

The modulatory role of BCL-2 gene in the regulation of apoptosis inHL-60 cells

published_or_final_versionPhysiologyDoctoralDoctor of Philosoph

Integrate CRISPR/Cas9 for protein expression of HLA-B*38:68Q via precise gene editing.

The determination of null- or low-expressed HLA alleles is clinically relevant in both hematopoietic stem cell transplantation and solid organ transplantation. We studied the expression level of a questionable (Q) HLA-B*38:68Q allele, which carries a 9-nucleotide (nt) deletion at codon 230-232 in exon 4 of HLA-B*38:01:01:01 using CRISPR/Cas9 gene editing technology. CRISPR/Cas9 gene editing of HLA-B*38:01:01:01 homozygous EBV B cell line resulted in one HLA-B*38:68Q/B*38:01:01:01 heterozygous and one HLA-B*38:68Q homozygous clone. Flow cytometric analysis of monoclonal anti-Bw4 antibody showed the protein expression of HLA-B*38:01:01:01 in homozygous cells was 2.2 fold higher than HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the expression of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 2.0 fold higher than HLA-B*38:68Q homozygous cells. The HLA-B*38:68Q expression was further confirmed using anti-B38 polyclonal antibody. Similarly, the expression of the HLA-B*38:01:01:01 homozygous cells was 1.5 fold higher than that of HLA-B*38:68Q/B*38:01:01:01 heterozygous cells, and the HLA-B*38:68Q/B*38:01:01:01 heterozygous cells was over 1.6 fold higher than that of HLA-B*38:68Q homozygous cells. The treatment of HLA-B*38:68Q homozygous cells with IFN-γ significantly increased its expression. In conclusion, we demonstrate that HLA-B*38:68Q is a low-expressing HLA allele. The CRISPR/Cas9 technology is a useful tool to induce precise gene editing in HLA genes to enable the characterization of HLA gene variants on expression and function