This blog is also published on Biocompare.
CRISPR genome editing has democratized gene editing, making it accessible to researchers across a wide array of fields. Plasmid-based CRISPR editing, one of the earliest methods, was a significant step forward due to relative ease of use and the ability to integrate all the necessary components for genome editing within a single vector.
However, the journey from the early plasmid-based methods to today's more advanced techniques has not been without challenges. The prolonged expression of CRISPR-associated proteins and guide RNAs from plasmids presented a major downside—extended exposure of the cells to these components increased the likelihood of off-target activity. This unintended editing could result in undesirable mutations, compromising the integrity of the genome and leading to potential downstream issues in functional studies or therapeutic applications.
Furthermore, plasmid-based methods often require antibiotic selection to ensure the expression of CRISPR components, which could affect cell viability and behavior, making it challenging to maintain the cells' original characteristics. These challenges were particularly pronounced for sensitive cell types like iPSCs or primary cells, as these cells are notoriously difficult to work with and are prone to stress-induced changes.
However, a recent market survey highlights that despite these drawbacks, a significant portion of researchers still rely on plasmid or lentivirus delivery methods, largely due to their familiarity and historical usage.
As CRISPR technology has evolved, the toolbox available to researchers has expanded significantly. One of the most notable advancements is the development of ribonucleoprotein (RNP) delivery systems. RNPs offer a more refined tool, allowing for transient expression of the CRISPR machinery, drastically reducing the risk of off-target effects across a wide range of cell types.
Fig 1. Pre-assembled ribonucleoprotein (RNP) complex creates a double-strand break. Unlike traditional plasmid-based approaches, RNP delivery of CRISPR-Cas9 components doesn’t require additional steps within the cell, like transcription and translation, and its transient presence within the cell minimizes the risk of off-target editing and downstream issues with post-editing cell functionality.
RNP delivery works by introducing a pre-assembled complex of Cas9 protein and a guide RNA directly into the cells. This approach ensures that the CRISPR components are only present in the cell for a short duration, minimizing the risk of off-target effects and reducing the potential for genomic instability. Moreover, RNPs do not rely on DNA intermediates and there is no need for antibiotic selection, preserving the natural state of the cells. This makes RNP delivery particularly attractive for applications in stem cell research, regenerative medicine, and other fields where maintaining cell integrity is crucial.
The case for outsourcing CRISPR editing
While new CRISPR methods like RNPs have opened doors for many researchers, the reality remains that not all laboratories have the resources, expertise, or time to fully utilize the growing CRISPR toolbox. A recent market survey underscores the challenges associated with do-it-yourself (DIY) CRISPR editing. Researchers spend an average of 61 hours on hands-on work per experiment, often facing the need to restart experiments multiple times before achieving success. This can lead to significant delays, with total project timelines extending up to 19 weeks and costs surpassing $18,000 per project.
Fig 2. Stepwise workflow for outsourcing CRISPR-edited cell pools and clones. A combination of intelligent bioinformatics, biological expertise, scalable automation, and strict QC processes are key features for ensuring efficient, consistent, and rapid cell engineering, even in the most complex and sensitive cell lines.
Even with access to the latest tools, the learning curve associated with mastering CRISPR techniques can be steep. For many researchers, the prospect of spending weeks or even months on a single experiment, only to achieve suboptimal results, is daunting. This is especially true in fast-paced research environments where meeting project deadlines is critical.
In this context, partnering with specialized CRISPR experts becomes an attractive option since they have spent time optimizing processes to deliver high-efficiency edits, reduce variability, and ensure the functionality of edited cells. Partnering with experts allows researchers to focus on their core objectives while leveraging the specialized capabilities of these partnerships.
Key considerations when choosing a partner
So how to choose a CRISPR editing partner?
Efficiency of the editing process: Efficiency is a crucial metric in CRISPR editing as it directly impacts the success of downstream applications. When choosing a partner, researchers should look for partners with a proven track record of delivering high editing efficiencies, particularly in challenging cell types. For instance, leveraging intelligent guide design strategies like multi-guide RNA CRISPR systems, ensures gene knockouts by deleting a fragment within an exon rather than relying on cuts from a single gRNA. Efficient editing methods are crucial when working with complex cell types like iPSCs or primary cells, where achieving efficient edits and maintaining cell functionality are paramount.
Consistency of the results: Consistency is equally important as variability in the editing process can lead to significant differences in experimental outcomes, making it difficult to draw reliable conclusions. A good partner will have well-established protocols that ensure consistent results across multiple edits and cell types.
Reducing time to results: Turnaround time is a critical factor in any research project. Delays in the CRISPR editing process can have a ripple effect on the entire project timeline, potentially leading to missed deadlines and increased costs. When partnering with a CRISPR expert, researchers should ensure that the team has the capacity to deliver results within the required timeframe, without compromising on quality.
The future of CRISPR editing
The democratization of CRISPR has transformed it from a niche technique requiring extensive expertise into a widely accessible tool that can be used across various scientific disciplines. As the technology continues to evolve, the future lies in leveraging advanced CRISPR platforms and partnering with experts who can meet the high efficiency, low variability, and robust post-edit functionality required.
Looking ahead, the continued development of CRISPR technology will likely focus on further increasing the precision and versatility of the tools available to researchers. Innovations such as base editing and prime editing, which allow for even more targeted and precise genetic modifications, are already beginning to make an impact in the field. These strategies will further expand the CRISPR toolbox, providing researchers with new options for tackling complex genetic challenges.
As CRISPR continues to advance, the importance of strategic partnerships in the field will only grow. By working with experts who can navigate the complexities of CRISPR editing and deliver high-quality results, researchers can focus on pushing the boundaries of science, confident that their genetic modifications will be accurate, efficient, and reliable. The future of CRISPR is bright, and with the right partnerships in place, the possibilities are endless.
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