METHODS FOR POLYNUCLEOTIDE INTEGRATION INTO THE GENOME OF BACILLUS USING DUAL CIRCULAR RECOMBINANT DNA CONSTRUCTS AND COMPOSITIONS THEREOF

Methods and compositions are provided for integrating genes of interest into the genome of a Bacillus sp. cell without the integration of a selectable marker into said genome. The methods employ a dual circular recombinant DNA system for introduction of a guide RNA/Cas endonuclease system (also refe...

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Bibliographic Details
Main Authors FRISCH, Ryan L, STUBBS, Stacey Irene Robida, SUH, Wonchul, ZIMMER, Derek Joseph
Format Patent
LanguageEnglish
French
Published 08.10.2020
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMBINATORIAL CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Summary:Methods and compositions are provided for integrating genes of interest into the genome of a Bacillus sp. cell without the integration of a selectable marker into said genome. The methods employ a dual circular recombinant DNA system for introduction of a guide RNA/Cas endonuclease system (also referred to as an RNA guided endonuclease, RGEN) as well as a donor DNA into a Bacillus sp. cell, and providing a highly effective system for inserting genes of interest into the genome of said Bacillus sp. cell. L'invention concerne des procédés et des compositions pour intégrer des gènes d'intérêt dans le génome d'une cellule de Bacillus sp . sans l'intégration d'un marqueur sélectionnable dans ledit génome. Les procédés utilisent un système d'ADN recombiné double circulaire pour l'introduction d'un système ARNg/endonucléase Cas (également appelé endonucléase guidée par ARN, RGEN) ainsi qu'un ADN donneur dans une cellule de Bacillus sp., et permettre d'obtenir un système hautement efficace pour insérer des gènes d'intérêt dans le génome de ladite cellule de Bacillus sp.
Bibliography:Application Number: WO2020US26503