Evidence map›Paper›PMID 31147565›Full record

ArticleScientific reports2019

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

Yuxin Yin, Elaine F Reed, Qiuheng Zhang

Abstract read
In one paragraph

Article in Scientific reports, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed
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1 · What the graph read from it

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2 · The registry

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3 · Its place in the literature

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4 · The record

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5 · Who and what money

Authors and funding

3 authors.

Yuxin YinUCLA Immunogenetics Center, Department of Pathology & Laboratory Medicine, Los Angeles, 90095, USA.
Elaine F ReedUCLA Immunogenetics Center, Department of Pathology & Laboratory Medicine, Los Angeles, 90095, USA.
Qiuheng ZhangUCLA Immunogenetics Center, Department of Pathology & Laboratory Medicine, Los Angeles, 90095, USA. jqzhang@mednet.ucla.edu.ORCID http://orcid.org/0000-0003-1439-6491

Funding

Regulation of Innate Immune Responses by Alloimmunity in Liver Ischemia-Reperfusion InjuryP01AI120944 · NIAID · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI Jerzy W Kupiec-Weglinski · 2017 to 2026
$20.0M
Project 3: Mapping the Evolution of Chronic Transplant Injury in the Context of CMV InfectionU19AI128913 · NIAID · UNIVERSITY OF CALIFORNIA LOS ANGELES · PI REED, ELAINE F, SARWAL, MINNIE M · 2017 to 2021
$11.2M
NIAID NIH HHS P01 AI120944NIAID NIH HHS U19 AI128913
6 · The paper itself

Abstract

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.

Indexed as

AllelesB-LymphocytesCell LineCRISPR-Cas SystemsExonsFeasibility StudiesGene EditingGene Expression RegulationHerpesvirus 4, HumanHeterozygoteHistocompatibility TestingHLA-B38 AntigenHomozygoteHumansPolymorphism, GeneticSequence DeletionHLA-B38 Antigen

Identifiers

PMID31147565
PMCPMC6542842

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.