Conventional genome editing approaches for engineering eukaryotic cells such as T-cells or plant cells using techniques like viral delivery can result in an exogenous gene or nucleic acid fragment being inserted at random locations and with multiple copies in the genome. These types of random insertions can, among other effects, disrupt the normal functioning of the genome.
However, the CRISPR/Cas9 system can be efficiently tailored for the specific placement of a gene or gene fragment within the genome. This approach to genome editing can result in better defined research tools, more effective gene and CAR T therapies, and greater efficiencies in plant trait expressions while preserving the normal functioning of the original genome.
MilliporeSigma (the Life Science business of Merck KGaA, Darmstadt, Germany) is a leader in developing innovative genome editing tools and technologies such as CRISPR and promoting its translation into genomic medicines to benefit patients and plant engineering to better feed the world’s population.
We are committed to making these technologies broadly available for research to help ensure that scientific development has the best chance of success.
This patent family covers successful integration of an external DNA sequence into the chromosome of eukaryotic cells using CRISPR.
MilliporeSigma, a leader in genome editing, has received several patents on its CRISPR-based eukaryotic genomic editing patent applications based upon International Application No. PCT/US2013/073307 in the following countries:
MilliporeSigma also has pending patent filings for its insertion CRISPR method in the U.S., Brazil, India, and Japan.
This invention covers ucleic acid integration using CRISPR Cas9 which can be used in gene editing for correction or disabling of diseased genes or adding genes for producing desired traits in plants and crops as well as many other applications. These fundamental method claims are necessary to achieve nucleic acid integration in eukaryotic cells. The IP is regarded as essential in conducting any homology directed repair that currently is a key gene editing approach.
The invention provides a method for integrating an exogenous nucleic acid sequence into a chromosomal sequence of a eukaryotic cell either in vivo, ex vivo or in vitro, as follows:
(i) at least one RNA-guided endonuclease comprising at least one nuclear localization signal or nucleic acid encoding at least one RNA-guided endonuclease comprising at least one nuclear localization signal, wherein the at least one RNA-guided endonuclease is a clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated (Cas) (CRISPR/Cas) type II system protein and the CRISPR/Cas type II system protein is a Cas9 protein, (ii) at least one guide RNA or DNA encoding at least one guide RNA, and (iii) at least one donor polynucleotide comprising the exogenous sequence; and b) Culturing the eukaryotic cell such that the guide RNA guides the RNA-guided endonuclease to a target site in the chromosomal sequence where the RNA-guided endonuclease introduces a double-stranded break, and the double-stranded break is repaired by a DNA repair process such that the exogenous sequence is integrated into the chromosomal sequence, wherein the method does not comprise a process for modifying the germ line genetic identity of a human being.
To enable access to this foundational scientific discovery, MilliporeSigma is offering non-exclusive licenses to this technology to academic and non-profit organizations, and commercial entities for internal research use.
In addition, non-exclusive, field-exclusive and disease or trait indication-exclusive commercial licenses are available for research, production, pharmaceutical and agricultural uses.
CRISPR research products purchased from MilliporeSigma will not require additional licenses unless the product is used outside the scope of the enclosed label license.
We look forward to hearing from you and welcome your interest in our IP and how it may assist you in achieving your goals. Please address all inquiries to:
Director of Business Development & Licensing
Gene Editing & Novel Modalities
A business of Merck KGaA, Darmstadt, Germany
Sigma-Aldrich Corp. | 2909 Laclede Avenue | Saint Louis, MO 63103 | USA
Phone: +1 703-740-6907
E-mail: firstname.lastname@example.org | www.milliporesigma.com