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10.1093/nar/gkw214 http://scihub22266oqcxt.onion/10.1093/nar/gkw214 C4857001!4857001!27036863     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Nucleic+Acids+Res 2016 ; 44 (8): 3801-10 Nephropedia Template TP {{tp|p=27036863|t=2016. CRISPR-mediated control of the bacterial initiation of replication |pdf=|usr=}}{{27036863}} gab.com Text CRISPR-mediated control of the bacterial initiation of replication JO: Nucleic Acids Res http://www.kidney.de/pget.php?&tw=1&pmid=27036863 #FOAvip Programmable control of the cell cycle has been shown to be a powerful tool in cell-biology studies. Here, we develop a novel system for controlling the bacterial cell cycle, based on binding of CRISPR/dCas9 to the origin-of-replication locus. Initiation of replication of bacterial chromosomes is accurately regulated by the DnaA protein, which promotes the unwinding of DNA at oriC. We demonstrate that the binding of CRISPR/dCas9 to any position within origin or replication blocks the initiation of replication. Serial-dilution plating, single-cell fluorescence microscopy, and flow-cytometry experiments show that ongoing rounds of chromosome replication are finished upon CRISPR/dCas9 binding, but no new rounds are initiated. Upon arrest, cells stay metabolically active and accumulate cell mass. We find that elevating the temperature from 37 to 42°C releases the CRISR/dCas9 replication inhibition, and we use this feature to recover cells from the arrest. Our simple and robust method of controlling the bacterial cell cycle is a useful asset for synthetic biology and DNA-replication studies in particular. The inactivation of CRISPR/dCas9 binding at elevated temperatures may furthermore be of wide interest for CRISPR/Cas9 applications in genomic engineering. Twit Text FOAVip CRISPR-mediated control of the bacterial initiation of replication ????Nucleic Acids Res http://www.kidney.de/pget.php?&tw=1&pmid=27036863 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=27036863 #FOAvip English Wikipedia <ref>{{Cite pmid|27036863}}</ref> CRISPR-mediated control of the bacterial initiation of replication #MMPMID27036863 Wiktor J; Lesterlin C; Sherratt DJ; Dekker C Nucleic Acids Res 2016[May]; 44 (8): 3801-10 PMID27036863show ga Programmable control of the cell cycle has been shown to be a powerful tool in cell-biology studies. Here, we develop a novel system for controlling the bacterial cell cycle, based on binding of CRISPR/dCas9 to the origin-of-replication locus. Initiation of replication of bacterial chromosomes is accurately regulated by the DnaA protein, which promotes the unwinding of DNA at oriC. We demonstrate that the binding of CRISPR/dCas9 to any position within origin or replication blocks the initiation of replication. Serial-dilution plating, single-cell fluorescence microscopy, and flow-cytometry experiments show that ongoing rounds of chromosome replication are finished upon CRISPR/dCas9 binding, but no new rounds are initiated. Upon arrest, cells stay metabolically active and accumulate cell mass. We find that elevating the temperature from 37 to 42°C releases the CRISR/dCas9 replication inhibition, and we use this feature to recover cells from the arrest. Our simple and robust method of controlling the bacterial cell cycle is a useful asset for synthetic biology and DNA-replication studies in particular. The inactivation of CRISPR/dCas9 binding at elevated temperatures may furthermore be of wide interest for CRISPR/Cas9 applications in genomic engineering. äCRISPR-mediated control of the bacterial initiation of replication (epmc).pdf DeepDyve Pubget Overpricing