T. Baba, T. Ara, M. Hasegawa, Y. Takai, Y. Okumura et al., Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection, Mol Syst Biol, vol.2, p.8, 2006.

J. Beal, T. Haddock-angelli, M. Gershater, K. De-mora, M. Lizarazo et al., Reproducibility of fluorescent expression from engineered biological constructs in E. coli, iGEM Interlab Study Contributors, vol.11, p.150182, 2016.

F. O. Bendezú, C. A. Hale, T. G. Bernhardt, and P. De-boer, RodZ (YfgA) is required for proper assembly of the MreB actin cytoskeleton and cell shape in E. coli, EMBO J, vol.28, pp.193-204, 2009.

D. Bikard, W. Jiang, P. Samai, A. Hochschild, F. Zhang et al., Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system, Nucleic Acids Res, vol.41, pp.7429-7437, 2013.

D. Bikard, C. W. Euler, W. Jiang, P. M. Nussenzweig, G. W. Goldberg et al., Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials, Nat Biotechnol, vol.32, pp.1146-1150, 2014.
URL : https://hal.archives-ouvertes.fr/hal-01103559

E. A. Boyle, J. Andreasson, L. M. Chircus, S. H. Sternberg, M. J. Wu et al., High-throughput biochemical profiling reveals sequence determinants of dCas9 off-target binding and unbinding, 2017.

, Proc Natl Acad Sci, vol.114, pp.5461-5466

A. M. Breier, H. Weier, and N. R. Cozzarelli, Independence of replisomes in Escherichia coli chromosomal replication, Proc Natl Acad Sci, vol.102, pp.3942-3947, 2005.

J. A. Brophy and C. A. Voigt, Antisense transcription as a tool to tune gene expression, Mol Syst Biol, vol.12, p.854, 2016.

T. Ça-gatay, M. Turcotte, M. B. Elowitz, J. Garcia-ojalvo, and G. M. Süel, Architecture-dependent noise discriminates functionally analogous differentiation circuits, Cell, vol.139, pp.512-522, 2009.

H. Cho, C. N. Wivagg, M. Kapoor, Z. Barry, P. Rohs et al., Bacterial cell wall biogenesis is mediated by SEDS and PBP polymerase families functioning semi-autonomously, Nat Microbiol, vol.1, p.16172, 2016.

L. Cui and D. Bikard, Consequences of Cas9 cleavage in the chromosome of Escherichia coli, Nucleic Acids Res, vol.44, pp.4243-4251, 2016.
URL : https://hal.archives-ouvertes.fr/pasteur-01967442

M. B. Elowitz, A. J. Levine, E. D. Siggia, and P. S. Swain, Stochastic gene expression in a single cell, Science, vol.297, pp.1183-1186, 2002.

I. Farasat and H. M. Salis, A biophysical model of CRISPR/Cas9 activity for rational design of genome editing and gene regulation, PLoS Comput Biol, vol.12, p.1004724, 2016.

D. G. Gibson, L. Young, R. Y. Chuang, J. C. Venter, C. A. Hutchison et al., Enzymatic assembly of DNA molecules up to several hundred kilobases, Nat Methods, vol.6, pp.343-345, 2009.

P. D. Hsu, E. S. Lander, and F. Zhang, Development and applications of CRISPRCas9 for genome engineering, Cell, vol.157, pp.1262-1278, 2014.

Y. Jiang, B. Chen, C. Duan, B. Sun, J. Yang et al., Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system, Appl Environ Microbiol, vol.81, pp.2506-2514, 2015.

D. L. Jones, P. Leroy, C. Unoson, D. Fange, V. Curi-c et al., , 2017.

, Kinetics of dCas9 target search in Escherichia coli, Science, vol.357, pp.1420-1424

D. Kennell and H. Riezman, Transcription and translation initiation frequencies of the Escherichia coli lac operon, J Mol Biol, vol.114, pp.1-21, 1977.

C. Kuscu, S. Arslan, R. Singh, J. Thorpe, and M. Adli, Genome-wide analysis reveals characteristics of off-target sites bound by the Cas9 endonuclease, Nat Biotechnol, vol.32, pp.677-683, 2014.

T. K. Lee, C. Tropini, J. Hsin, S. M. Desmarais, T. S. Ursell et al., A dynamically assembled cell wall synthesis machinery buffers cell growth, Proc Natl Acad Sci, vol.111, pp.4554-4559, 2014.

X. Li, Y. Jun, M. J. Erickstad, S. D. Brown, A. Parks et al., ) tCRISPRi: tunable and reversible, one-step control of gene expression, Sci Rep, vol.6, p.39076, 2016.

A. J. Meeske, E. P. Riley, W. P. Robins, T. Uehara, J. J. Mekalanos et al., SEDS proteins are a widespread family of bacterial cell wall polymerases, Nature, vol.537, pp.634-638, 2016.

A. A. Nielsen and C. A. Voigt, Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks, Mol Syst Biol, vol.10, p.763, 2014.
DOI : 10.15252/msb.20145735

URL : http://msb.embopress.org/content/10/11/763.full.pdf

G. S. Pall and A. J. Hamilton, Improved northern blot method for enhanced detection of small RNA, Nat Protoc, vol.3, pp.1077-1084, 2008.

J. M. Peters, A. Colavin, H. Shi, T. L. Czarny, M. H. Larson et al., A comprehensive, CRISPR-based functional analysis of essential genes in bacteria, Cell, vol.165, pp.1493-1506, 2016.

G. Pósfai, V. Kolisnychenko, Z. Bereczki, and F. R. Blattner, Markerless gene replacement in Escherichia coli stimulated by a double-strand break in the chromosome, Nucleic Acids Res, vol.27, pp.4409-4415, 1999.

L. S. Qi, M. H. Larson, L. A. Gilbert, J. A. Doudna, J. S. Weissman et al., Repurposing CRISPR as an RNA-guided platform for sequencespecific control of gene expression, Cell, vol.152, pp.1173-1183, 2013.

E. Sauvage, F. Kerff, M. Terrak, J. A. Ayala, and P. Charlier, The penicillin-binding proteins: structure and role in peptidoglycan biosynthesis, FEMS Microbiol Rev, vol.32, pp.234-258, 2008.

A. Shafferman and D. R. Helinski, Structural properties of the beta origin of replication of plasmid R6K, J Biol Chem, vol.258, pp.4083-4090, 1983.

L. So, A. Ghosh, C. Zong, L. A. Sepúlveda, R. Segev et al., General properties of transcriptional time series in Escherichia coli, Nat Genet, vol.43, pp.554-560, 2011.

S. H. Sternberg, S. Redding, M. Jinek, E. C. Greene, and J. A. Doudna, DNA interrogation by the CRISPR RNA-guided endonuclease Cas9, Nature, vol.507, pp.62-67, 2014.
DOI : 10.1016/j.bpj.2013.11.3848

URL : https://doi.org/10.1016/j.bpj.2013.11.3848

F. St-pierre, L. Cui, D. G. Priest, D. Endy, I. B. Dodd et al., One-step cloning and chromosomal integration of DNA, ACS Synth Biol, vol.2, pp.537-541, 2013.
DOI : 10.1021/sb400021j

F. V. Subach, G. H. Patterson, S. Manley, J. M. Gillette, J. Lippincott-schwartz et al., Photoactivatable mCherry for high-resolution twocolor fluorescence microscopy, Nat Methods, vol.6, pp.153-159, 2009.
DOI : 10.1038/nmeth.1298

URL : http://europepmc.org/articles/pmc2901231?pdf=render

M. D. Szczelkun, M. S. Tikhomirova, T. Sinkunas, G. Gasiunas, T. Karvelis et al., Direct observation of R-loop formation by single RNA-guided Cas9 and Cascade effector complexes, Proc Natl Acad Sci, vol.111, pp.9798-9803, 2014.

Y. Taniguchi, P. J. Choi, G. Li, H. Chen, M. Babu et al., Quantifying E. coli proteome and transcriptome with single-molecule sensitivity in single cells, Science, vol.329, pp.533-538, 2010.

T. Ursell, T. K. Lee, D. Shiomi, H. Shi, C. Tropini et al., Rapid, precise quantification of bacterial cellular dimensions across a genomicscale knockout library, BMC Biol, vol.15, p.17, 2017.

A. J. Waite, N. W. Frankel, Y. S. Dufour, J. F. Johnston, J. Long et al., Nongenetic diversity modulates population performance, Mol Syst Biol, vol.12, p.895, 2016.
DOI : 10.15252/msb.20178149

URL : http://msb.embopress.org/content/msb/14/1/e8149.full.pdf

J. Wiktor, C. Lesterlin, D. J. Sherratt, and C. Dekker, CRISPR-mediated control of the bacterial initiation of replication, Nucleic Acids Res, vol.44, pp.3801-3810, 2016.

X. Wu, D. A. Scott, A. J. Kriz, A. C. Chiu, P. D. Hsu et al., Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells, Nat Biotechnol, vol.32, pp.670-676, 2014.

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