References
1. Barrangou R, Fremaux C, Deveau H,
Richards M, Boyaval P, Moineau S, Romero
DA, Horvath P (2007) CRISPR provides
acquired resistance against viruses in prokaryotes. Science 315:1709–1712
2. Garneau JE, Dupuis M, Villion M, Romero
DA, Barrangou R, Boyaval P, Fremaux C,
Horvath P, Magada ´n AH, Moineau S (2010)
The CRISPR/Cas bacterial immune system
cleaves bacteriophage and plasmid DNA.
Nature 468:67–71
3. Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang
S (2015) Multigene editing in the Escherichia
coli genome via the CRISPR-Cas9 system.
Appl Environ Microbiol 81:2506–2514
4. Shapiro RS, Chavez A, Collins JJ (2018)
CRISPR-based genomic tools for the manipulation of genetically intractable microorganisms. Nat Rev Microbiol 16:333–339
5. Cobb RE, Wang Y, Zhao H (2015) Highefficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/
Cas system. ACS Synth Biol 4:723–728
6. DiCarlo JE, Norville JE, Mali P, Rios X, Aach J,
Church GM (2013) Genome engineering in
Saccharomyces cerevisiae using CRISPR-Cas
systems. Nucleic Acids Res 41:4336–4343
7. Shan Q, Wang Y, Li J, Zhang Y, Chen K,
Liang Z, Zhang K, Liu J, Xi JJ, Qiu JL, Gao
C (2013) Targeted genome modification of
crop plants using a CRISPR-Cas system. Nat
Biotechnol 31:686–688
8. Matthews LA, Simmons LA (2014) Bacterial
nonhomologous end joining requires teamwork. J Bacteriol 196:3363–3365
9. Weterings E, van Gent DC (2004) The mechanism of non-homologous end-joining: a synopsis of synapsis. DNA Repair (Amst)
3:1425–1435
10. Bikard D, Hatoum-Aslan A, Mucida D, Marraffini LA (2012) CRISPR interference can
prevent natural transformation and virulence
acquisition during in vivo bacterial infection.
Cell Host Microbe 12:177–186
11. Cui L, Bikard D (2016) Consequences of Cas9
cleavage in the chromosome of Escherichia coli.
Nucleic Acids Res 44:4243–4251
12. Fernandes LGV, Guaman LP, Vasconcellos SA,
Heinemann MB, Picardeau M, Nascimento
ALTO (2019) Gene silencing based on
RNA-guided catalytically inactive Cas9
(dCas9): a new tool for genetic engineering in
Leptospira. Sci Rep 9:1839
13. Qi LS, Larson MH, Gilbert LA, Doudna JA,
Weissman JS, Arkin AP, Lim WA (2013)
Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene
expression. Cell 152:1173–1183
14. Demarre G, Gue ´rout AM, MatsumotoMashimo C, Rowe-Magnus DA, Marlie `re P,
Mazel D (2005) A new family of mobilizable
suicide plasmids based on broad host range
R388 plasmid (IncW) and RP4 plasmid
(IncPalpha) conjugative machineries and their
cognate Escherichia coli host strains. Res Microbiol 156:245–255
15. Pappas CJ, Benaroudj N, Picardeau M (2015)
A replicative plasmid vector allows efficient
complementation of pathogenic Leptospira
strains.
Appl
Environ
Microbiol
81:3176–3181
16. Zhukova A, Fernandes LG, Hugon P, Pappas
CJ, Sismeiro O, Coppe ´e JY, Becavin C,
Malabat C, Eshghi A, Zhang JJ, Yang FX,
Picardeau M (2017) Genome-wide transcriptional start site mapping and sRNA identification in the pathogen. Front Cell Infect
Microbiol 7:10
17. Picardeau M (2008) Conjugative transfer
between Escherichia coli and Leptospira spp. as
a new genetic tool. Appl Environ Microbiol
74:319–322
122
Luis Guilherme Virgı ´lio Fernandes and Ana Lucia Tabet Oller Nascimento
1. Barrangou R, Fremaux C, Deveau H,
Richards M, Boyaval P, Moineau S, Romero
DA, Horvath P (2007) CRISPR provides
acquired resistance against viruses in prokaryotes. Science 315:1709–1712
2. Garneau JE, Dupuis M, Villion M, Romero
DA, Barrangou R, Boyaval P, Fremaux C,
Horvath P, Magada ´n AH, Moineau S (2010)
The CRISPR/Cas bacterial immune system
cleaves bacteriophage and plasmid DNA.
Nature 468:67–71
3. Jiang Y, Chen B, Duan C, Sun B, Yang J, Yang
S (2015) Multigene editing in the Escherichia
coli genome via the CRISPR-Cas9 system.
Appl Environ Microbiol 81:2506–2514
4. Shapiro RS, Chavez A, Collins JJ (2018)
CRISPR-based genomic tools for the manipulation of genetically intractable microorganisms. Nat Rev Microbiol 16:333–339
5. Cobb RE, Wang Y, Zhao H (2015) Highefficiency multiplex genome editing of Streptomyces species using an engineered CRISPR/
Cas system. ACS Synth Biol 4:723–728
6. DiCarlo JE, Norville JE, Mali P, Rios X, Aach J,
Church GM (2013) Genome engineering in
Saccharomyces cerevisiae using CRISPR-Cas
systems. Nucleic Acids Res 41:4336–4343
7. Shan Q, Wang Y, Li J, Zhang Y, Chen K,
Liang Z, Zhang K, Liu J, Xi JJ, Qiu JL, Gao
C (2013) Targeted genome modification of
crop plants using a CRISPR-Cas system. Nat
Biotechnol 31:686–688
8. Matthews LA, Simmons LA (2014) Bacterial
nonhomologous end joining requires teamwork. J Bacteriol 196:3363–3365
9. Weterings E, van Gent DC (2004) The mechanism of non-homologous end-joining: a synopsis of synapsis. DNA Repair (Amst)
3:1425–1435
10. Bikard D, Hatoum-Aslan A, Mucida D, Marraffini LA (2012) CRISPR interference can
prevent natural transformation and virulence
acquisition during in vivo bacterial infection.
Cell Host Microbe 12:177–186
11. Cui L, Bikard D (2016) Consequences of Cas9
cleavage in the chromosome of Escherichia coli.
Nucleic Acids Res 44:4243–4251
12. Fernandes LGV, Guaman LP, Vasconcellos SA,
Heinemann MB, Picardeau M, Nascimento
ALTO (2019) Gene silencing based on
RNA-guided catalytically inactive Cas9
(dCas9): a new tool for genetic engineering in
Leptospira. Sci Rep 9:1839
13. Qi LS, Larson MH, Gilbert LA, Doudna JA,
Weissman JS, Arkin AP, Lim WA (2013)
Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene
expression. Cell 152:1173–1183
14. Demarre G, Gue ´rout AM, MatsumotoMashimo C, Rowe-Magnus DA, Marlie `re P,
Mazel D (2005) A new family of mobilizable
suicide plasmids based on broad host range
R388 plasmid (IncW) and RP4 plasmid
(IncPalpha) conjugative machineries and their
cognate Escherichia coli host strains. Res Microbiol 156:245–255
15. Pappas CJ, Benaroudj N, Picardeau M (2015)
A replicative plasmid vector allows efficient
complementation of pathogenic Leptospira
strains.
Appl
Environ
Microbiol
81:3176–3181
16. Zhukova A, Fernandes LG, Hugon P, Pappas
CJ, Sismeiro O, Coppe ´e JY, Becavin C,
Malabat C, Eshghi A, Zhang JJ, Yang FX,
Picardeau M (2017) Genome-wide transcriptional start site mapping and sRNA identification in the pathogen. Front Cell Infect
Microbiol 7:10
17. Picardeau M (2008) Conjugative transfer
between Escherichia coli and Leptospira spp. as
a new genetic tool. Appl Environ Microbiol
74:319–322
122
Luis Guilherme Virgı ´lio Fernandes and Ana Lucia Tabet Oller Nascimento