[McGovern Institute for Brain Research at MIT]

Mammoth Biosciences—the CRISPR-based diagnostics developer whose co-founders include genome editing pioneer Jennifer Doudna, Ph.D.—said today it has achieved a key milestone toward being first to market a CRISPR-based disease detection platform by exclusively licensing two U.S. patents granted to the regents of the University of California that cover CRISPR collateral cleavage diagnostic systems.

U.S. Patent No. 10,253,365 and U.S. Patent No. 10,337,051  cover comprehensive techniques that will enable Mammoth to offer DNA and RNA detection diagnostics that apply CRISPR gene editing. Patent ‘365 covers Type V CRISPR/Cas effector proteins for cleaving single stranded DNAs (ssDNAs) and detecting target DNAs, while patent ‘051 covers methods and compositions for detecting a single-stranded target RNA.

“Essentially, collateral cleavage serves as an amplification and a readout for saying the CRISPR protein successfully identified whatever nucleic acid you told it to identify,” Mammoth Biosciences CEO and co-founder Trevor Martin, Ph.D., told Clinical OMICs. “This technique gives you exquisite sensitivity and specificity like the best molecular techniques, but you’re getting it in a multiplexable, isothermal, very easy-to-use format.”

“The specificity is an especially great aspect of the CRISPR system,” Martin added. “You can even do really specific single SNP analysis using this system, while still maintaining a great level of sensitivity, and on top of that, carry out multiplexing.”

Martin and Doudna, of UC Berkeley, co-founded Mammoth with Lucas Harrington, Mammoth’s Chief Discovery Officer; Janice Chen, Ph.D., the company’s Chief Research Officer; and Ashley Tehranchi, Ph.D., who served as CTO until May.

Doudna, Harrington, and Chen are listed at co-inventors of Patent ‘365, along with Enbo Ma, Senior Staff Scientist in Doudna’s lab.

On Patent ‘051, Doudna is listed among co-inventors, which include three former graduate students in her lab—Mitchell R. O’Connell, Ph.D., now a principal investigator at University of Rochester (NY) Medical Center; Alexandra East-Seletsky, PhD, now a scientist with Arrakis Therapeutics, and Spencer Charles Knight, Ph.D.—as well as Doudna’s husband Jamie H.D. Cate, Ph.D., Professor of Chemistry, Biochemistry and Molecular Biology at UC Berkeley.

Detecting Multiple Conditions

Mammoth Biosciences says its easy-to-use and affordable point-of-care test will enable fast, simultaneous detection of multiple conditions, in real-time, both in the hospital and at home, through a single credit card-sized strip form factor. [Mammoth Biosciences]
Mammoth’s co-founders brought the company from stealth mode in April 2018, disclosing

plans to develop an easy-to-use and affordable point-of-care test enabling fast, simultaneous detection of multiple conditions, in real-time, both in the hospital and at home, through a single credit card-sized strip form factor.

Once at-home users complete the test, Mammoth envisions them securely uploading an anonymous photo of the strip to the company’s corresponding application for remote analysis. According to the company, users can receive their confidential results and professional advice, all in potentially less than 15 minutes, without having to collect samples and mail them for analysis.

A year ago, Mammoth raised $23 million dollars to fund the initial development of the test. The diagnostics apply CRISPR to search and find DNA or RNA that indicate the presence of disease, akin to how Google searches for information on the Web

Mammoth’s platform publicly uses three unique CRISPR proteins: Cas12, which targets double-stranded DNA; Cas13, which targets and recognizes single-stranded RNA; and Cas14, which targets single-stranded DNA. Used together, the proteins enable the diagnostic to sense virtually any type of nucleic acid.

All three are programmable by a guide RNA protein that matches up with the sequence users want to detect. Then the protein cleaves reporter molecules to confirm a match has been found.

“You first bioinformatically design a guide RNA for those targets. The guide RNAs are going to be specific to the proteins and target, and that’s how you program the CRISPR proteins to identify malaria versus Ebola or something else,” Martin said.

Once Cas12, Cas13 or Cas14 finds its matching target, the protein can cleave the reporter molecules, for example, producing a color change that indicates the presence of a specific DNA or RNA sequence.

“Democratize Disease Detection”

Mammoth Biosciences staff

Mammoth has committed itself to “democratize disease detection” by developing easy-to-use, affordable, and effective tests usable in a variety of formats from central labs and point-of-care settings, such as hospitals and clinics, to even home use.

“In a lab setting, you can have a different level of targets you’re going after. In a consumer setting, you’d want it in a very simple, easy to use form factor and most importantly, that’s easy to understand,” Martin said. “At Mammoth, we’re most focused on delivering information that’s actually actionable.”

Mammoth is among at least two companies founded by CRISPR pioneers focused on developing diagnostics using the gene editing technology. In March, Feng Zhang, Ph.D., of The Broad Institute of MIT and Harvard was among co-founders who launched Sherlock Biosciences. Sherlock has said it plans to leverage synthetic biology and intellectual property related to the CRISPR patents held by the Broad and Harvard University’s Wyss Institute.

Sherlock takes its name from one of its two core technologies—Specific High-sensitivity Enzymatic Reporter unLOCKing (SHERLOCK). The other core technology is dubbed INSPECTR (INternal Splint-Pairing Expression Cassette Translation Reaction), a molecular diagnostic platform using synthetic-biology developed in the Wyss Institute lab of company co-founder James J. Collins, Ph.D.

“At Mammoth, we think competition is great, and it‘s just further validation of how exciting this technology is,” Martin said. “Our patents are really a great validation of Mammoth being at the forefront of this field.”

Martin said Mammoth is looking at applications for its diagnostic technology that go beyond healthcare. The company, he said, is developing tests and teaming up with diagnostic partners to explore applications in industries that include agriculture, oil and gas, and forensics.

Mammoth is not disclosing its partners or its pricing for its diagnostics, though Martin did say the company is looking to increase its accessibility to consumers.

“We’re really trying to make it so that you don’t always have to go necessarily to the doctor’s office to get molecular information about your health,” Martin added. “We can be a catalyst for the rest of the healthcare system to actually be delivered to the point of care.”

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