CRISPR-Cas9 attacking HIV in an immune cell

CRISPR's Next Frontier: Engineering Immunity for the Future

Reese Marlowe in Health & Wellness December 2025 • 4 min read.

"How a novel cellular platform is optimizing CRISPR/Cas9 technology for AIDS gene therapy, bringing hope and new strategies to combat HIV."

In the ongoing battle against HIV and AIDS, scientists are constantly seeking new and innovative therapeutic strategies. Despite the advancements in antiretroviral therapies, AIDS remains a significant global health challenge. The rise of drug-resistant HIV strains underscores the urgent need for more effective, long-term solutions.

One promising avenue of research involves gene therapy, which aims to modify a patient's own cells to combat the virus. Immortalized cell lines, particularly those representing the major targets of HIV in humans—macrophages and CD4+ T lymphocytes—are invaluable for this research. These cell lines allow scientists to study HIV infection and test new therapeutic approaches in a controlled laboratory setting.

Recently, researchers have focused on using CRISPR/Cas9 technology to engineer HIV-resistant cells. This gene-editing tool allows for precise modifications to a cell's DNA, offering the potential to disrupt viral entry or replication. By optimizing CRISPR/Cas9 technology within a novel cellular platform, scientists are paving the way for more effective and targeted gene therapies for AIDS.

Engineering HIV Resistance: How CRISPR/Cas9 Targets CCR5

CRISPR-Cas9 attacking HIV in an immune cell

A critical aspect of HIV infection involves the virus's ability to bind to specific receptors on the surface of immune cells. The co-receptors CCR5 and CXCR4 are essential for HIV to enter CD4+ T cells. Disrupting these co-receptors can prevent the virus from infecting cells, providing a potential therapeutic strategy. Many T cell lines do not express adequate levels of CCR5, limiting their utility for HIV research.

To address this limitation, researchers have developed a novel cellular platform using CRISPR/Cas9 technology to create a CCR5-expressing CD4+ T cell line. This involves inserting the CAG promoter into the CCR5 gene's promoter region via homologous recombination. This process induces the expression of CCR5 from its endogenous locus, resulting in a cell line that is highly permissive to HIV infection. Here are the steps taken:

Selecting the Right Cell Line: Jurkat T cells were chosen for their high expression levels of CD4 and CXCR4 genes.
CRISPR/Cas9 Knock-In: The ubiquitous CAG promoter was inserted into the CCR5 gene promoter to enhance CCR5 expression.
Verification of Recombination: PCR and Southern blotting were used to confirm homologous recombination events and rule out random integration.
Creation of Jurkat-KI-R5: The resulting cell line, Jurkat-KI-R5, stably expresses high levels of CCR5.

The resulting Jurkat-KI-R5 cell line has proven to be highly susceptible to CCR5-tropic HIV infection. Unlike parental Jurkat cells, Jurkat-KI-R5 cells produce detectable levels of p24 antigen, a marker of HIV replication. This novel cellular platform provides a valuable tool for HIV research and the development of gene therapies targeting AIDS.

Future Directions: Eradicating HIV with Precision Gene Editing

The development of Jurkat-KI-R5 cells opens up new avenues for gene therapy strategies aimed at disrupting CCR5. By using CRISPR/Cas9 technology to target the CCR5 gene in these cells, researchers can generate HIV-resistant CD4+ T cells. These modified cells can then be used to study the efficacy of different gene-editing approaches and optimize protocols for clinical applications.

About this Article -

This article was crafted using a human-AI hybrid and collaborative approach. AI assisted our team with initial drafting, research insights, identifying key questions, and image generation. Our human editors guided topic selection, defined the angle, structured the content, ensured factual accuracy and relevance, refined the tone, and conducted thorough editing to deliver helpful, high-quality information.See our About page for more information.

This article is based on research published under:

DOI-LINK: 10.1007/s13238-017-0453-z, Alternate LINK

Title: Using A Novel Cellular Platform To Optimize Crispr/Cas9 Technology For The Gene Therapy Of Aids

Subject: Cell Biology

Journal: Protein & Cell

Publisher: Springer Science and Business Media LLC

Authors: Jingjin He, Thanutra Zhang, Xuemei Fu

Published: 2017-08-18

Everything You Need To Know

What is CRISPR/Cas9 technology, and how is it being used to combat HIV?

CRISPR/Cas9 technology is a gene-editing tool that allows scientists to make precise modifications to a cell's DNA. In the context of HIV, researchers are using CRISPR/Cas9 to engineer HIV-resistant cells. This involves targeting specific genes, like CCR5, that HIV uses to infect cells. By disrupting these genes, scientists aim to prevent the virus from entering and replicating within immune cells, offering a potential long-term therapeutic strategy against AIDS.

What is the role of CCR5 in HIV infection, and why is it a target for gene therapy?

CCR5 is a co-receptor on the surface of CD4+ T cells that HIV uses to enter and infect these cells. HIV relies on CCR5 to bind to the cell, allowing the virus to inject its genetic material. Disrupting CCR5 prevents HIV from infecting CD4+ T cells, which are a major target for HIV. The novel cellular platform developed uses CRISPR/Cas9 technology to create a CCR5-expressing CD4+ T cell line, making these cells highly susceptible to HIV infection. This allows researchers to study the effects of disrupting CCR5 and develop targeted gene therapies.

How was the Jurkat-KI-R5 cell line created, and what makes it useful for HIV research?

The Jurkat-KI-R5 cell line was created using CRISPR/Cas9 technology to enhance CCR5 expression in Jurkat T cells. The process involved inserting the CAG promoter into the CCR5 gene promoter to increase CCR5 levels. This was achieved through homologous recombination, confirmed by PCR and Southern blotting. The resulting Jurkat-KI-R5 cell line stably expresses high levels of CCR5 and is highly susceptible to HIV infection. This makes it an invaluable tool because it allows scientists to study HIV infection and test new therapeutic approaches in a controlled laboratory setting, accelerating the development of gene therapies for AIDS.

What are the key steps involved in using CRISPR/Cas9 to engineer HIV-resistant cells in the novel cellular platform?

The key steps in the novel cellular platform to engineer HIV-resistant cells involve: selecting the right cell line, specifically Jurkat T cells because of the high expression levels of CD4 and CXCR4 genes; CRISPR/Cas9 knock-in, inserting the CAG promoter into the CCR5 gene promoter to enhance CCR5 expression, verification of recombination using PCR and Southern blotting to confirm homologous recombination events and rule out random integration; and finally the creation of Jurkat-KI-R5, a cell line that stably expresses high levels of CCR5. This process enables researchers to study how gene editing approaches can disrupt the CCR5 gene, leading to the development of HIV-resistant CD4+ T cells for potential clinical applications.

What are the future implications of this research for the treatment of AIDS, and what are the next steps?

The development of the Jurkat-KI-R5 cell line opens up new avenues for gene therapy strategies that aim to eradicate HIV. The next steps involve using CRISPR/Cas9 technology to target the CCR5 gene in the Jurkat-KI-R5 cells, creating HIV-resistant CD4+ T cells. These modified cells will then be used to study the efficacy of different gene-editing approaches and optimize protocols for clinical applications. Ultimately, the goal is to develop effective gene therapies that can provide long-term solutions, potentially eradicating HIV and enhancing immunity, moving beyond the limitations of current antiretroviral therapies.