Modalities

Gene Editing

We are initially pointing our platform towards CRISPR and gene editing, as we believe there is a significant and persistent unmet need that AI is uniquely positioned to address.

The transformation of microbial CRISPR-based enzymes like SpCas9 into gene editing tools has spurred a revolution in genetic medicine. However, current technologies still fall short in many key areas, limiting the promise that gene editing has to dramatically improve human health. Indeed, gene editing systems that are mixed and matched with components lifted from nature often have functional tradeoffs that significantly limit their reach. For example, PAM relaxation expands edit flexibility at the expense of precision, complex gene writing systems expand the scope of potential edits at the expense of efficiency, and large integration systems offer the promise of whole gene insertion but struggle with deliverability.

Our ability to read and write proteins from scratch offers an opportunity to reimagine gene editing systems from the ground up, breaking free of evolutionary constraints or labor-intensive, single-attribute protein engineering approaches. In stark contrast to current approaches, our AI platform enables us to start with final applications and work backward to custom-design solutions. This includes everything from precisely tuning existing CRISPR scaffolds to creating entirely novel editing systems.

OpenCRISPR™

Profluent is committed to advancing innovation and development in the gene editing community, to bring new treatments to patients with major unmet needs. Under the OpenCRISPR initiative, we are releasing the world’s first open-source, AI-generated gene editor. With this launch, Profluent demonstrates the first successful precision editing of the human genome with customizable gene editors designed from scratch with AI.

Through the OpenCRISPR training process, Profluent’s large language models (LLMs) learned from a massive scale of protein sequences overlaid with biological context to generate millions of diverse CRISPR-like proteins that do not occur in nature, thereby exponentially expanding virtually all known CRISPR families.

We have decided to launch OpenCRISPR-1 (one of our AI-created gene editors) as an initial open-source release, making it freely available to license for ethical research and commercial uses. OpenCRISPR-1 maintains the prototypical architecture of a Type II Cas9 nuclease but is >400 mutations away from SpCas9 and nearly 200 mutations away from any other known natural CRISPR-associated protein. We also used our LLMs to generate a synthetic guide RNA that could be assembled with OpenCRISPR-1.

In our recently released manuscript, we describe both the generation and characterization of OpenCRISPR-1. When delivered via plasmids in HEK293T cells, OpenCRISPR-1 showed comparable on-target editing efficiency and higher specificity relative to SpCas9, across a wide range of genomic on- and off-targets. Furthermore, when combined with AI-generated deaminases, OpenCRISPR-1 was able to assume functionality of a base editing architecture, showing robust A-to-G editing at a panel of target sites.

Studies are ongoing to examine genome-wide specificity and behavior of OpenCRISPR-1 as a purified Ribonucleoprotein (RNP) complex. OpenCRISPR-1 is our first release. We encourage folks to use it, test it, and provide feedback so we can iterate and advance the initiative forward.

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Frequently Asked Questions

Reach out to info@profluent.bio

OpenCRISPR is interesting, but I have more needs; what does Profluent offer?

We are open to collaboratively iterate and customize an AI-designed solution that is a perfect match for your specific therapeutic application. This ranges from customized gene editors, antibodies, and broader enzymes. Please email partnerships@profluent.bio.

Is there a way to share my experience using OpenCRISPR with Profluent?

We expressly welcome any feedback on OpenCRISPR and especially sharing of any observations as you’re using the system. If you find that certain attributes could be changed or improved for your particular needs, please reach out!

Do you provide protocols?

Please see our pre-print in Biorxiv link for a general protocol in addition to a readme protocol that accompanies the sequence release. Other general protocols for editing enzymes should also be compatible.

Will there be additional OpenCRISPR releases in the future?

Stay tuned...

What does the license include?

The current release includes the protein sequence of OpenCRISPR-1 along with a compatible AI-generated gRNA, though it is also compatible with canonical Cas9 gRNAs.

If OpenCRISPR is truly open source, then why do I need to sign a license agreement?

The sequence is freely available via the pre-print. We considered many factors to make accessing OpenCRISPR as frictionless and lightweight as possible; chief among these was ensuring its ethical and safe use. For this reason, if OpenCRISPR users wish to use the molecule for commercial therapeutic uses, we require them to execute a simple license agreement that includes obligations to use the tool for ethical purposes only, in addition to other terms of use.

Are you really not asking for anything?

In addition to abiding by our terms of use, we kindly ask that you allow us to acknowledge you as a user and to let us know when any products using OpenCRISPR advance to the clinic or commercial stages.

Have you filed IP on OpenCRISPR?

Yes.

Why are you releasing OpenCRISPR™ free of charge – what’s the catch?

There is no catch. OpenCRISPR is free for commercial use to any users who take a license. In a world where gene editing technologies can be difficult to access for both researchers and patients for various reasons, we felt the need to put our company mission into action and release some of the byproducts of our prolific protein design engine to enable more discoveries in the gene editing industry. For partners where further customization and expanded features for OpenCRISPR or another system might be desired, we offer a high-touch collaboration model.

What is OpenCRISPR-1?

OpenCRISPR-1 is an AI-created gene editor, consisting of a Cas9-like protein and guide RNA, fully developed using Profluent’s large language models (LLMs). The OpenCRISPR-1 protein maintains the prototypical architecture of a Type II Cas9 nuclease but is hundreds of mutations away from SpCas9 or any other known natural CRISPR-associated protein. You can view OpenCRISPR-1 as a drop-in replacement for many protocols that need a Cas9-like protein with an NGG PAM and you can even use it with canonical SpCas9 gRNAs. OpenCRISPR-1 can be fused in a deactivated or nickase format for next generation gene editing techniques like base, prime, or epigenome editing. Find out more in our preprint.