Difference between revisions of "Part:BBa K4621081"

 
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<partinfo>BBa_K4621081 short</partinfo>
 
<partinfo>BBa_K4621081 short</partinfo>
  
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This CRISPRi fragment contains four parts : BBa _ K4621070, BBa _ K3875019, BBa _ K4621031, and BBa _ K3081104, covering the dCas9 and sgRNA necessary for the CRISPRi system. The CRISPRi system is suitable for a variety of Streptomyces.[1] In this study, it was used to inhibit the expression of GQS52 _ 08040 gene in SCUT-3 to produce more ectoine and hydroxyectoine.
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This CRISPRi fragment contains the dCas9 and sgRNA necessary for the CRISPRi system. The CRISPRi system is suitable for a variety of Streptomyces.[1] In this study, it was used to inhibit the expression of GQS52 _ 08040 gene in SCUT-3 to produce more ectoine and hydroxyectoine.
  
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Usage, Biology and Characterization
 
  
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===Usage and Biology===
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https://static.igem.wiki/teams/4621/wiki/parts/mechanism-of-crispri.png
  
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Due to the limited types of plasmids available for SCUT-3, we inserted the CRISPRi fragment into the previously constructed plasmid by homologous recombination to achieve simultaneous stable expression of multiple target genes. Subsequently, we verified the effectiveness of the CRISPRi system in the fermentation of ordinary LB, high-salt LB and shrimp shells.
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Fig.1 CRISPRi system used in the study
  
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Due to the limited types of plasmids available for SCUT-3, we inserted the CRISPRi-488 fragment into the previously constructed plasmid by homologous recombination to achieve simultaneous stable expression of multiple target genes. Subsequently, we verified the effectiveness of the CRISPRi-488 system (Pi8 or ProCi8 in the figure below) in the fermentation of ordinary LB, high-salt LB and shrimp shells.
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https://static.igem.wiki/teams/4621/wiki/parts/fermentation-under-high-salt-environment.png
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Fig.2 Performance test on CRISPRi strains
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https://static.igem.wiki/teams/4621/wiki/parts/fermentation-using-shrimp-shells-p.png
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Fig. 3 Ectoine/hydroxyectoine production using shrimp shell waste of CRISPRi strains
  
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===Usage and Biology===
 
  
 
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Revision as of 12:14, 10 October 2023


CRISPRi-488

This CRISPRi fragment contains the dCas9 and sgRNA necessary for the CRISPRi system. The CRISPRi system is suitable for a variety of Streptomyces.[1] In this study, it was used to inhibit the expression of GQS52 _ 08040 gene in SCUT-3 to produce more ectoine and hydroxyectoine.


Usage and Biology

mechanism-of-crispri.png

Fig.1 CRISPRi system used in the study

Due to the limited types of plasmids available for SCUT-3, we inserted the CRISPRi-488 fragment into the previously constructed plasmid by homologous recombination to achieve simultaneous stable expression of multiple target genes. Subsequently, we verified the effectiveness of the CRISPRi-488 system (Pi8 or ProCi8 in the figure below) in the fermentation of ordinary LB, high-salt LB and shrimp shells.

fermentation-under-high-salt-environment.png Fig.2 Performance test on CRISPRi strains

fermentation-using-shrimp-shells-p.png Fig. 3 Ectoine/hydroxyectoine production using shrimp shell waste of CRISPRi strains


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 30
    Illegal SpeI site found at 4549
    Illegal PstI site found at 18
    Illegal PstI site found at 2443
    Illegal PstI site found at 2677
    Illegal PstI site found at 3889
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 4526
    Illegal SpeI site found at 4549
    Illegal PstI site found at 18
    Illegal PstI site found at 2443
    Illegal PstI site found at 2677
    Illegal PstI site found at 3889
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 482
    Illegal BglII site found at 1277
    Illegal BglII site found at 4046
    Illegal BamHI site found at 222
    Illegal BamHI site found at 1571
    Illegal BamHI site found at 4514
    Illegal BamHI site found at 4719
    Illegal XhoI site found at 895
    Illegal XhoI site found at 3241
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 30
    Illegal SpeI site found at 4549
    Illegal PstI site found at 18
    Illegal PstI site found at 2443
    Illegal PstI site found at 2677
    Illegal PstI site found at 3889
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 30
    Illegal SpeI site found at 4549
    Illegal PstI site found at 18
    Illegal PstI site found at 2443
    Illegal PstI site found at 2677
    Illegal PstI site found at 3889
    Illegal NgoMIV site found at 1839
    Illegal NgoMIV site found at 2210
    Illegal NgoMIV site found at 2413
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3421
    Illegal BsaI.rc site found at 4300
    Illegal SapI site found at 813
    Illegal SapI site found at 2103
    Illegal SapI.rc site found at 3300