Difference between revisions of "Part:BBa K2325003"

Revision as of 07:48, 22 October 2017

T7 phage N20-sgRNA

The BBa_K2325003 is a DNA sequence contains N20 associated with sgRNA. The N20-1 is selected from the genome of T7 phage, which is responsible for the synthesis of the DNA packaging protein during phage infection. This part confers resistance against T7 phage upon a strain containing the Cas9 and sgRNA parts. This part can be considered as an improvement of the BBa_K2325003, as they contain same N20 sequence but different sgRNA sequence. We fused crRNA and tracrRNA as a single synthetic guide RNA (sgRNA), while UBC2013 constructed crRNA (repeat-spacer array) and tracrRNA separately. In their construct, the crRNA and tracrRNA were separately transcript, followed by the combination of these transcripts to form the sgRNA. The resist efficiency may decrease when the crRNA and tracrRNA not 100% match successfully. The way we constructed the sgRNA obviated the need for processing the transcribed CRISPR array (pre-crRNA) into individual crRNA components, helping to promote the function of resistance against T7 phage.

Sequence and Features

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]

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 The BBa_K2325003 is a DNA sequence contains N20 associated with sgRNA. The N20-1 is selected from the genome of T7 phage, which is responsible for the synthesis of the DNA packaging protein during phage infection.
 This part confers resistance against T7 phage upon a strain containing the Cas9 and sgRNA parts.
-This part can be considered as an improvement of the (BBa_K1129012), as they contain same N20 sequence but different sgRNA sequence. We fused crRNA and tracrRNA as a single synthetic guide RNA (sgRNA), which obviated the need for processing the transcribed CRISPR array (pre-crRNA) into individual crRNA components.
+<b>This part can be considered as an improvement of the <partinfo>BBa_K2325003</partinfo></b>, as they contain same N20 sequence but different sgRNA sequence.  We fused crRNA and tracrRNA as a single synthetic guide RNA (sgRNA), while UBC2013 constructed crRNA (repeat-spacer array) and tracrRNA separately. In their construct, the crRNA and tracrRNA were separately transcript, followed by the combination of these transcripts to form the sgRNA. The resist efficiency may decrease when the crRNA and tracrRNA not 100% match successfully. The way we constructed the sgRNA obviated the need for processing the transcribed CRISPR array (pre-crRNA) into individual crRNA components, helping to promote the function of resistance against T7 phage.
-The way we constructed the part also promoted the function of resistance against T7 phage.
 <!-- Add more about the biology of this part here