CRISPR-Cas9 Mediated Capsule Gene Silencing in Escherichia coli
Materials and Methods: We suppressed genes involved in capsule expression of E.coli by CRISPR cas9 process. The constructed E.coli was confirmed by microscopic smear, transmission electron microscopy and T7 phage influence assay.
Results: The results were shown that the inhibition of capsule production was carried out successfully and there was not any capsule layer around the bacteria.
Conclusion: E. coli without any capsule around may proper for replacement of it with other molecules in future.
Esvelt KM, Wang HH. Genome-scale engineering for systems and synthetic biology. Mol Syst Biol. 2013;9:641.
Wang HH, Church GM. Multiplexed genome engineering and genotyping methods applications for synthetic biology and metabolic engineering. Methods Enzymol. 2011;498:409-26.
Deltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao Y, Pirzada ZA, Eckert MR, Vogel J, Charpentier E. CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature. 2011;471:602-7.
Mao F, Dam P, Chou J, Olman V, Xu Y. Door: a database for prokaryotic operons, Nucleic Acids Res. 2009;37: 459-63.
Mark P Stevens, Bradley R Clarke, Ian S. Roberts. Regulation of the Escherichia coli K5 capsule gene cluster by transcription antitermination. Mol Microbiol. 1997;24(5):1001-12.
Díaz-Visurraga J, Cárdenas G, García A. Morphological changes induced in bacteria as evaluated by electron microscopy. Micros: Sci, Technol, Appl and Educ. 2010;1:307-15.
Scholl D, Adhya S, Merril C, Escherichia coli K1 s Capsule Is a Barrier to Bacteriophage T7. Appl and Environ Microbiol. 2005;71(8):4872-4.
Enyeart P, Chirieleison S, Dao M, Perutka J, Quandt E, Yao J, et al. Generalized bacterial genome editing using mobile group II introns and Cre‐lox. Mol Sys Biol. 2013;9:685
Cao J, Wu L, Zhang Sh. An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting. Nucleic Acids Research 2016;44(19):2506-14.
Jiang Y, Chen B, Duan Ch, Sun B, Yang J, Yang Sh. Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System. Applied and Environmental Microbiology 2015;81(7):2506-15.
Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA. RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013;31:233-9.
Gonzalez-Garcia V, Bocanegra R, Pulido-Cid M, Martin-Benito J, Cuervo A. Characterization of the initial steps in the T7 DNA ejection process. Bacteriophage 2015;5(3):e1056904-1.