Difference between revisions of "Part:BBa K3971003"
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<partinfo>BBa_K3971003 short</partinfo> | <partinfo>BBa_K3971003 short</partinfo> | ||
| + | ===Usage and Biology=== | ||
This sequence is a gRNA target that is downstream of a TTC 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]. | This sequence is a gRNA target that is downstream of a TTC 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]. | ||
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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]. | 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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Revision as of 17:26, 9 August 2021
gRNA target for Cpf1 based editing in the Neutral Site 1 of Synechococcus elongatus UTEX 2973.
Usage and Biology
This sequence is a gRNA target that is downstream of a TTC 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 13
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]