Difference between revisions of "Part:BBa K1062003:Design"

 
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<partinfo>BBa_K1062003 short</partinfo>
 
<partinfo>BBa_K1062003 short</partinfo>
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<partinfo>BBa_K1062003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1062003 SequenceAndFeatures</partinfo>
  
  
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===Design Notes===
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=='''Design Notes'''==
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Part:BBa_K1062003 is a gRNA that specifically targets the end of the XylE gene.
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==='''gRNA'''===
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A gRNA is a 102 nt sequence in which contains: a 20 nt sequence which will contain the sequence of the intended target, a 42 nt Cas9-binding hairping, and a 40 nt transcription terminator.
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[[File:gRNA with NNN sequence.png]]
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'''Figure 1: An image show a general picture of a gRNA.'''
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An important thing to note when using or creating a gRNA is that before the 20nt sequence, '''''there must be a 3nt sequence called a PAM sequence which contain this sequence 5'-NGG-3''''''. This 3nt sequence is what the Cas9 use as a binding signal. It is also important to point out that the PAM sequence is never in the gRNA itself.
  
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[[File:pam.png]]
  
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===Source===
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'''Figure 2: An example of a PAM sequence before the 20nt target sequence.'''
  
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==Sources==
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The gRNA was design by the iGEM team by using APE file. Once the gRNA was finalized, the team ordered primers and used PCR sewing in order to create the gRNA.
  
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===References===
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==References==

Latest revision as of 20:44, 17 September 2013

Guide RNA (gRNA) target for XylE (end of gene)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Part:BBa_K1062003 is a gRNA that specifically targets the end of the XylE gene.

gRNA

A gRNA is a 102 nt sequence in which contains: a 20 nt sequence which will contain the sequence of the intended target, a 42 nt Cas9-binding hairping, and a 40 nt transcription terminator.

GRNA with NNN sequence.png

Figure 1: An image show a general picture of a gRNA.


An important thing to note when using or creating a gRNA is that before the 20nt sequence, there must be a 3nt sequence called a PAM sequence which contain this sequence 5'-NGG-3'. This 3nt sequence is what the Cas9 use as a binding signal. It is also important to point out that the PAM sequence is never in the gRNA itself.

Pam.png

Figure 2: An example of a PAM sequence before the 20nt target sequence.

Sources

The gRNA was design by the iGEM team by using APE file. Once the gRNA was finalized, the team ordered primers and used PCR sewing in order to create the gRNA.

References