淋巴细胞和Jurkat细胞系是开发CRISPR-Cas9方法的有用替代品,以纠正白细胞粘附缺陷基因组缺陷
Ahmad R Ramadan1,2, Noureddine Ben Khalaf3, Khaled Trabelsi4
1Laboratory of Transmission, Control and Immunobiology of Infections (LR16IPT02), Institut Pasteur de Tunis, University of Tunis El Manar, Tunis, Tunisia.
Frontiers in bioengineering and biotechnology
|April 7, 2025
概括
通过CRISPR-Cas9基因编辑,在白细胞粘附性缺陷1型 (LAD1) 患者细胞中成功恢复了CD18的表达. 这种精确的校正通过使免疫功能稳定,为LAD1提供了潜在的治疗方法.
科学领域:
- 遗传学和基因组学 遗传学和基因组学
- 免疫学 免疫学 免疫学
- 生物技术是生物技术.
背景情况:
- 白细胞粘附性缺陷1型 (LAD1) 是一种严重的遗传性免疫疾病,由ITGB2基因突变引起.
- 缺乏CD18/β-2整合素会损害白细胞粘附和免疫反应,导致复发性感染.
- CRISPR-Cas9基因编辑为纠正LAD1.1中的遗传缺陷提供了潜在的治疗策略.
研究的目的:
- 开发和验证CRISPR-Cas9基因编辑策略,以纠正LAD1.1中的ITGB2突变.
- 评估患者衍生细胞中基因校正的效率和特异性.
- 评估CD18表达的恢复及其潜在的治疗影响.
主要方法:
- 设计并测试了ITGB2基因的CRISPR-Cas9介导裂变的三个导向RNA (gRNA).
- 在LAD1患者的淋巴细胞细胞中使用了一种高效的gRNA,具有增强的Cas9 (espCas9) 和同质导向修复 (HDR) 模板.
- 通过流细胞计量量化CD18表达,并使用全基因组测序 (WGS) 评估非目标效应.
主要成果:
- 在初步测试中,2-rev gRNA显示出最高的裂变效率.
- 在使用优化的CRISPR-Cas9系统与HDR-2的LAD1细胞中,CD18表达的恢复率达到了23%.
- 全基因组测序证实了高特异性,没有可检测的非目标突变或不必要的插入.
结论:
- CRISPR-Cas9基因编辑是一种精确有效的方法,用于纠正LAD1.1.中的ITGB2突变.
- 达到的CD18表达水平足以可能将疾病表型从严重转变为中度.
- 这种方法对临床转化有希望,特别是在CD34+造血干细胞疗法中,为潜在的长期治愈提供了希望.
相关概念视频
Lymphoid Cells and Tissues
627
Lymphoid cells and tissues are integral to the immune system, which is crucial in maintaining our body's defense against harmful pathogens. They form the building blocks of lymphoid organs, which include the spleen, thymus, and lymph nodes.
Lymphoid cells consist of various types of immune system cells. These include B and T lymphocytes, which are responsible for producing antibodies and killing infected cells, respectively. Dendritic cells act as messengers between the innate and adaptive...
Lymphoid cells consist of various types of immune system cells. These include B and T lymphocytes, which are responsible for producing antibodies and killing infected cells, respectively. Dendritic cells act as messengers between the innate and adaptive...
627
Cells of the Adaptive Immune Response
646
The T and B lymphocytes of the adaptive immune system develop from common lymphoid progenitor cells in the bone marrow. These progenitors give rise to precursors that eventually develop into both T and B lymphocytes. As these precursors mature, they gain the ability to detect and respond to foreign antigens in the body, a process known as immunocompetence. Additionally, these precursors acquire self-tolerance, a process that ensures they do not react to self-antigens. This intricate system...
646
Secondary Lymphoid Organs
825
Secondary organs, including lymph nodes, the spleen, and mucosa-associated lymphoid tissue (MALT), work harmoniously to protect us from disease and infection.
The spleen is a vital organ in the lymphatic system, nestled in the upper left side of the abdomen. It is composed of two primary regions: the red pulp and the white pulp, each having distinct functions. The red pulp performs a significant role in blood filtration. It efficiently purges the blood of old or damaged red blood cells and...
The spleen is a vital organ in the lymphatic system, nestled in the upper left side of the abdomen. It is composed of two primary regions: the red pulp and the white pulp, each having distinct functions. The red pulp performs a significant role in blood filtration. It efficiently purges the blood of old or damaged red blood cells and...
825
Primary Lymphoid Organs
963
Primary lymphoid organs are pivotal in the formation, development, and maturation of lymphocytes, the white blood cells that serve as the backbone of our immune system. This crucial function underscores their fundamental role in maintaining our overall health and immunity. The two primary lymphoid organs of prime importance are the red bone marrow and the thymus.
The red bone marrow is a soft, spongy tissue nestled in the interior of long bones such as the humerus and femur. It is the site...
The red bone marrow is a soft, spongy tissue nestled in the interior of long bones such as the humerus and femur. It is the site...
963
Classification of Leukocytes
1.3K
Leukocytes are classified into two groups based on the presence or absence of cytoplasmic granules. Granular leukocytes, which contain granules, belong to the myeloid lineage and are divided into three subtypes: neutrophils, eosinophils, and basophils. These cells are roughly spherical and characterized by the granules in their cytoplasm.
Neutrophils are the most abundant type of granular leukocytes, comprising 50-70% of all leukocytes. They feature small, evenly distributed granules and a...
Neutrophils are the most abundant type of granular leukocytes, comprising 50-70% of all leukocytes. They feature small, evenly distributed granules and a...
1.3K
B Cell Activation and Differentiation
1.3K
The adaptive immune response, a sophisticated defense mechanism, relies on the activation and differentiation of B lymphocytes, or B cells. These processes enable our bodies to mount a tailored response against specific pathogens such as bacteria, free virus particles, toxins, and parasites.
When naive B cells encounter a specific antigen that can bind to the B cell receptor (BCR) on their surface, they undergo sensitization to respond to the antigen's presence. Sensitization begins with...
When naive B cells encounter a specific antigen that can bind to the B cell receptor (BCR) on their surface, they undergo sensitization to respond to the antigen's presence. Sensitization begins with...
1.3K


