The Rockefeller University

Institut Pasteur

Eligo Bioscience

The Rockfeller University

Massachusetts Institute of Technology

Institut National De La Sante Et De La Recherche Medicale

The Broad Institute, Inc.

Assistance Publique-hopitaux De Paris

Universite Paris Cité

Fondation Imagine

Université Paris Descartes

David Bikard

Luciano Marraffini

Jesus Fernandez Rodriguez

Xavier Duportet

Antoine Decrulle

Lun Cui

Feng Zhang

Philippe J Sansonetti

Neville Espi Sanjana

David Benjamin Turitz Cox

Gerard Eberl

Wenyan Jiang

Valerie Gaboriau-Routhiau

Robert Heller

Nadine Cerf Bensussan

Pamela Schnupf

Sébastien Baur

Alexander Hawkins-Hooker

Gérard Eberl

Valérie Gaboriau-Routhiau

Provided are compositions and methods for selectively reducing the amount of antibiotic resistant and/or virulent bacteria in a mixed bacteria population, or for reducing any other type of unwanted bacteria in a mixed bacteria population. The compositions and methods involve targeting bacteria that are differentiated from other members of the population by at least one unique clustered regularly interspaced short palindromic repeats (CRISPR) targeted DNA sequence. The compositions and methods can be readily adapted to target any bacteria or any bacteria plasmid, or both.

Sequence specific antimicrobials

The invention relates to the improvement of endonuclease-based antimicrobials by blocking DNA repair of double-strand break(s) (DSB(s)) in prokaryotic cells. In this respect, the invention especially concerns a method involving blocking DNA repair after a nucleic acid has been submitted to DSB, in particular by a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) associated programmable double-strand endonuclease. The invention particularly relates to the use of an exogenous molecule that inhibits DNA repair, preferably a protein that binds to the ends of the double-stranded break to block DSB repair. The invention also relates to vectors, particularly phagemids and plasmids, comprising nucleic acids encoding nucleases and Gam proteins, and a pharmaceutical composition and a product containing these vectors and their application.

Sequence-specific antimicrobials by blocking DNA repair

The present invention relates to a vector, preferably included in a delivery vehicle, comprising no more than 100, preferably no more than 10, restriction sites recognized by the restriction enzymes encoded by each bacterium of a group of bacteria of interest. The invention also relates to the use of said vector, preferably included in a delivery vehicle, as a drug, especially in the treatment of a disease in a patient in need thereof.

Optimized vector for delivery in microbial populations

The present invention relates to a vector, preferably included in a delivery vehicle, comprising no more than (100), preferably no more than (10), restriction sites recognized by the restriction enzymes encoded by each bacterium of a group of bacteria of interest. The invention also relates to the use of said vector, preferably included in a delivery vehicle, as a drug, especially in the treatment of a disease in a patient in need thereof.

The present invention relates to an in vitro method of culturing a segmented filamentous bacterium strain, comprising co-culturing said segmented filamentous bacterium strain with a eukaryotic host cell, wherein the culture is performed at an Olevel inferior to 5% in a rich tissue culture liquid medium containing bacterial medium components including iron. The present invention also relates to methods for genetically modifying a segmented filamentous bacterium strain comprising a step a culturing the strain in vitro.

Method of culturing segmented filamentous bacteria in vitro

Provided are Cas9 enzymes that have mutations that enhance their properties, relative to un-mutated Cas9. The altered Cas9 enzymes exhibit i) an increased rate of spacer acquisition, or ii) increased cleavage efficiency of targets with NAG PAMs, or a combination of i) and ii). The altered Cas9 enzymes comprise an amino acid substitution of 1473 and K500 in aor similar Cas9 enzyme. Also provided are polynucleotides, including expression vectors that encode the Cas9 enzymes, cells that contain the polynucleotides, and methods of making and using such cells. The disclosure includes tagging, or labelling bacteria, and for enhancing phage acquired immunity in bacteria, such as those used in industrial processes, including the food and beverage industry, such as the dairy industry. The food products are also included.

Compositions and methods using Cas9 with enhanced spacer acquisition function

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Paris, France

David Bikard