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Lab+Life Scientist June/July 2024

Page 12

gene editing iStock.com/Artem_Egorov

CRISPR-Cas gene editing eliminates HIV in lab

New research presented at this year’s European Congress of Clinical Microbiology and Infectious Diseases (ECCMID 2024), held in Barcelona from 27–30 April, shows how the latest CRISPR-Cas gene editing technology can be used to eliminate all traces of HIV from infected cells in the laboratory. The studies, conducted at Amsterdam University Medical Center (Amsterdam UMC), present a significant breakthrough in the search for a cure for HIV.

C

The scientists deployed CRISPR-Cas

successful, enhancing its delivery to HIV-infected

molecular scissors and two gRNAs against

cells. Moreover, they were able to target ‘hidden’

RISPR-Cas gene editing technology

‘conserved’ HIV sequences, meaning they focused

HIV reservoir cells by focusing on specific

is a groundbreaking technique that allows for

on parts of the virus genome that stay the same

proteins found on the surfaces of these cells

precise alterations to the genomes of living

across all known HIV strains, and achieved cure

(CD4+ and CD32a+).

organisms, enabling scientists to accurately target

of HIV-infected T cells. By focusing on these

“We have developed an efficient combinatorial

and modify specific segments of an organism’s

conserved sections, the approach aims to provide

CRISPR-attack on the HIV virus in various

DNA. Functioning like molecular ‘scissors’ with

a broad-spectrum therapy capable of combating

cells and the locations where it can be hidden in

the guidance of guide RNA (gRNA), CRISPR-

multiple HIV variants effectively.

reservoirs, and demonstrated that therapeutics can

Cas can cut the DNA at designated spots. This

However, the team found that the size of the

be specifically delivered to the cells of interest,” the

action facilitates either the deletion of unwanted

vehicle or ‘vector’ used to transport the cassette

scientists stated. They said their findings represent

genes or the introduction of new genetic material

encoding the therapeutic CRISPR-Cas reagents

a pivotal advancement towards designing a cure

into an organism’s cells, paving the way for

into the cells presented logistical challenges,

strategy for HIV, while acknowledging that their

advanced therapies.

as it was too large. Thus, they trialled various

work at this stage is only a proof of concept.

One of the significant challenges in HIV

techniques to reduce the size of the cassette — and

“Our next steps involve optimising the

treatment is the virus’s ability to integrate its

therefore the vector system itself. In simpler terms,

delivery route to target the majority of the HIV

genome into the host’s DNA, making it extremely

they’re attempting to pack oversized luggage

reservoir cells,” the researchers said. “We will

difficult to eliminate. Numerous potent antiviral

into a compact car for a journey to the infected

combine the CRISPR therapeutics and receptor-

drugs are currently in use for treating HIV

cell, leading them to find ways to downsize the

targeting reagents and move to preclinical

infection — but despite their efficacy, lifelong

‘luggage’ (cassette) for easier transport. Another

models to study in detail the efficacy and safety

antiviral therapy is essential, as HIV can rebound

issue they wanted to overcome was reaching the

aspects of a combined cure strategy. This will be

from established reservoirs when treatment is

HIV reservoir cells that ‘rebound’ when HIV

instrumental to achieving preferential CRISPR-

halted. CRISPR-Cas genome editing thus provides

antiretroviral treatment is stopped.

Cas delivery to the reservoir cells and avoiding

The researchers further evaluated various

delivery into non-reservoir cells. This strategy

“Our aim is to develop a robust and safe

CRISPR-Cas systems from different bacteria

is to make this system as safe as possible for

combinatorial CRISPR-Cas regimen, striving for

to determine their effectiveness and safety in

future clinical applications.

an inclusive ‘HIV cure for all’ that can inactivate

treating CD4+ T cells infected with HIV. They

“We hope to achieve the right balance between

diverse HIV strains across various cellular

shared results from two systems, saCas9 and

efficacy and safety of this cure strategy. Only

contexts,” the Amsterdam team said. With HIV

cjCas. SaCas9 showed outstanding antiviral

then can we consider clinical trials of ‘cure’ in

able to infect different types of cells and tissues in

performance, managing to completely inactivate

humans to disable the HIV reservoir. While these

the body, each with its own unique environment

HIV with a single guide RNA (gRNA) and

preliminary findings are very encouraging, it is

and characteristics, the researchers are searching

excise (cut out) the viral DNA with two gRNAs.

premature to declare that there is a functional

for a way to target HIV in all of these situations.

The strategy of minimising the vector size was

HIV cure on the horizon.”

a new means to target HIV DNA.

12 This issue is sponsored by | AMS Instrumentation & Calibration Solutions | www.ams-ic.com.au


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