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CRISPR Gene Editing Best Practices and the Importance of Applying Caution

Researchers and scientists all over the world perform CRISPR (clustered regularly interspaced short palindromic repeat) gene editing to edit the DNA in cells, whether belonging to a human, plant, bacteria, or virus. Not only is this gene editing approach simple and efficient, but it enables scientists to target multiple genes simultaneously and easily customize target DNA.

That said, CRISPR gene editing isn’t without limitations. Not only can CRISPR techniques affect regions of DNA outside the target, but ethical issues can arise from gene editing, especially in studies involving human embryos. As a result, the medical research community must continue to develop CRISPR gene editing techniques and the regulations surrounding these.

As progression takes place in this space, many researchers follow the life sciences journal BioTechniques to stay afloat of developments in CRISPR gene editing. This peer-reviewed journal explores the latest in gene editing, noting why scientists must uphold caution to ensure responsible, safe practice.

Here, we’ll delve into three top CRISPR gene editing practices that BioTechniques recommends, why researchers most apply caution to the process, and the direction we can expect the CRISPR industry to take.

Top CRISPR Gene Editing Practices

Scientists should follow various practices while performing CRISPR gene editing. In particular, BioTechniques highlights the following as three of the most important practices.

First, scientists should opt for the best CRISPR gene editing method for the research that they are conducting. They can choose from a variety of methods that suit different cell lines in a variety of medical and scientific fields. Some systems are better suited to specific cell and tissue types than others.

Second, scientists should employ the technologies and software that best support the CRISPR technique they will use. There are several technologies available, each offering different advantages, making them appropriate for different cell types.

Third, scientists should make ethical decisions surrounding their use of CRISPR. They must perform gene editing in line with regulations, which is especially important in research that involves human embryos. While regulations usually ban this kind of research, studies that involve human subjects may be deemed ethical if the benefits of gene editing outweigh the downsides. In this situation, scientists must uphold the utmost transparency and meet all legal requirements during the gene editing process.

Applying Caution to CRISPR Gene Editing

Challenges can arise during the CRISPR gene editing process, so scientists should apply caution throughout this process. These challenges typically surround effectiveness, unexpected mutations, and ethics.

Effectiveness

As the tools needed to measure how effective CRISPR gene editing is are often costly, these tools aren’t always accessible to scientists. That’s why, historically, scientists only measured the efficacy of CRISPR in the places where they had created edits. Now, scientists must often interrogate an entire genome, a process that requires modern tools and typically generates a large volume of data for analysis.

Ethics

The ethical use of CRISPR gene editing is a hotly contested area among scientists, particularly when it comes to human embryos. Overall, many scientists support the use of CRISPR in experiments that involve human embryos where there is a pressing medical need for gene editing. In these cases, research findings could fuel developments in diagnoses of and treatments for genetic disorders that currently have limited testing and treatment routes.

However, the effects of CRISPR gene editing vary depending on the mutation, which can make the process unpredictable. Although there is evidence that CRISPR is safe in some applications, there isn’t yet much advanced research that demonstrates the process is safe for human embryos.

BioTechniques has conducted a study to unpack researchers’ opinions on the ethical implementation of CRISPR gene editing. The study concluded that 63% of respondents supported the concept of human embryo experiments where the embryos won’t be brought to term, 55% supported the concept of preventing serious genetic disorders in human embryos that will be brought to term, 9% supported the concept of enhancing traits in human embryos (making edits without a medical need to do so), and 18% disagreed with CRISPR practices in any form.

Unexpected Mutations

If DNA undergoes unexpected mutations, or if scientists don’t modify DNA correctly, the CRISPR process can be unreliable. While scientists have examined unwanted alterations, they haven’t yet understood the full consequences of how the mutations play out in different organisms. For example, while some studies report adverse effects of CRISPR/Cas9 that are associated with pronounced on-target structural variants and off-target mutations, others contradict these findings.

One example of unexpected mutations in a CRISPR study is the study that Uppsala University and SciLifeLab’s National Genomics Infrastructure undertook in 2022. During this study, scientists analyzed the genomes of more than 1,000 zebrafish from two generations, demonstrating that the CRISPR/Cas9 method can inadvertently cause mutations in DNA, which future generations can inherit.

Mutations occurred in the wrong genomic locations, and some instances reported DNA fragments that were undergoing changes and were bigger than expected. Overall, the insertions and deletions caused structural variations that led to 6% of outcomes in founder larvae across both generations. As a result, the scientists highlighted the importance of thorough validation and careful consideration when deciding whether to use CRISPR/Cas9 for medical purposes.

The health care industry is already seeing scientists develop counteractive processes for genes that are associated with specific cell and tissue types. Although these treatments don’t pose a risk of intergenerational aberration, scientists should take note of the need for caution that these findings demonstrate.

Future Developments in CRISPR Gene Editing

Although scientists haven’t yet fully optimized the safety and efficacy of CRISPR, they have taken large steps forward in CRISPR gene editing technologies. They can apply these technologies to determine the effectiveness of a CRISPR technique and highlight any faults in the method. Meanwhile, the latest algorithms make CRISPR gene editing more time efficient and make it easier to identify and edit proteins.

Given that CRISPR is one of the quickest-evolving technology fields and acts as both an editing and detection tool, the gene editing approach has fast become a solution in many applications. As the gene editing technique continues to advance, scientists are likely to utilize CRISPR to:

  •       Perform advanced screening to interrogate genomes and determine genetic changes.
  •       Target heart disease, obesity, and leukemia.
  •       Identify whether patients have specific viruses and, by extension, make quick diagnoses.
  •       Team CRISPR technologies with other technologies to progress precision editing.
  •       Further the agriculture and plant production industry by enabling the production of healthier food products and allowing farmers to breed higher proportions of female cattle and chickens for milk and eggs.

BioTechniques is an open-access journal that publishes the latest in lab methods and instrumentation for scientists and researchers who specialize in the life sciences and related disciplines. This way, these professionals can stay atop fast-changing techniques and technologies in western blotting, next-generation sequencing (NGS), chromatography, polymerase chain reaction (PCR), and CRISPR gene editing. They can also visit BioTechniques’ multimedia website to access additional articles, interviews, podcasts, eBooks, webinars, videos, and key industry discussions involving the growing global research community.

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