Difference between revisions of "Part:BBa K3971029"

Revision as of 18:07, 20 October 2021

gRNA target for Cpf1 based editing in the Neutral Site 3 of Synechococcus elongatus UTEX 2973.

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Usage and Biology

This sequence is a gRNA target that is downstream of a TTG PAM sequence for Cpf1. This gRNA target sequence is to be cloned into the pSL2680 (Addgene Plasmid #85581) plasmid to make modifications in the Neutral site 1 of Synechococcus elongatus UTEX 2973, which it targets [1].

Neutral sites are loci in the genome with unknown functions and that can be disrupted without affecting cellular viability or causing any distinct phenotypes [2]. Cpf1 also known as Cas12a, can be used to make targeted modifications of DNA or RNA and is small in size, creating staggered instead of blunt cuts. It does not require tracrRNA and needs a 42-44 nucleotide crRNA. It also cuts distal to the PAM site without disrupting the target sequence and thus allows for future modifications at the same site [3].

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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 <partinfo>BBa_K3971029 short</partinfo>
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+-- Add more about the biology of this part here
 ===Usage and Biology===
+This sequence is a gRNA target that is downstream of a TTG PAM sequence for Cpf1. This gRNA target sequence is to be cloned into the pSL2680 (Addgene Plasmid #85581) plasmid to make modifications in the Neutral site 1 of Synechococcus elongatus UTEX 2973, which it targets [1].
+Neutral sites are loci in the genome with unknown functions and that can be disrupted without affecting cellular viability or causing any distinct phenotypes [2]. Cpf1 also known as Cas12a, can be used to make targeted modifications of DNA or RNA and is small in size, creating staggered instead of blunt cuts. It does not require tracrRNA and needs a 42-44 nucleotide crRNA. It also cuts distal to the PAM site without disrupting the target sequence and thus allows for future modifications at the same site [3].
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