Favorite Parts
Nluc-dCas9 fusion protein
(BBa_K1689010)
Nluc was fused to the N terminus of dCas9 to form the Nluc-dCas9 fusion protein. It is used to bind a DNA sequence guided by sgRNA. Paired with Cluc-dCas9 fusion protein (BBa_K1689009) and sgRNAs, our paired dCas9 reporter system were constructed.
Cluc-dCas9 fusion protein
(BBa_K1689009)
Cluc was fused to the N terminus of dCas9 to form the Cluc-dCas9 fusion protein. It is used to bind a DNA sequence guided by sgRNA. Paired with Nluc-dCas9 fusion protein (BBa_K1689010) and sgRNAs, our paired dCas9 reporter system were constructed.
sgRNA generator
(BBa_K1689000)
sgRNA generator is used for generating sgRNA which recognizes targeted DNA, then guiding dCas9 to it.
Part List
| Favorite | Name | Type | Description | Design |
|---|---|---|---|---|
| √ | BBa_K1689000 | Composite | sgRNA generator | ZHAO Xuejin |
| √ | BBa_K1689008 | Composite | Coding sequence of dCas9-Nluc | ZHANG Yihao |
| √ | BBa_K1689009 | Composite | Coding sequence Cluc-dCas9 | ZHANG Yihao |
| BBa_K1689010 | Compostie | Coding sequence of N-luc-dCas9 | ZHANG Yihao | |
| BBa_K1689001 | Coding | Coding sequence of STV-N-luc | ZHANG Yihao | |
| BBa_K1689002 | Coding | Coding sequence of Cluc-STV | ZHANG Yihao | |
| BBa_K1689003 | Coding | Coding sequence of Nluc416-FRB | Xiang Li | |
| BBa_K1689004 | Coding | Coding sequence of Nluc398-FRB | Xiang Li | |
| BBa_K1689005 | Coding | Coding sequence of FKBP-Cluc398 | Xiang Li | |
| BBa_K1689006 | Coding | Coding sequence of FKBP-Cluc394 | Xiang Li | |
| BBa_K1689007 | Composite | Coding sequence of dCas9-C-luc | ZHANG Yihao | |
| BBa_K1689011 | Composite | delta alpha segment of lacz fused with dCas9, His tag allows for protein purification | Jiaofeng Li | |
| BBa_K1689012 | Composite | N terminal of β-lactamase fused with dCas9 | Jiaofeng Li | |
| BBa_K1689013 | Compostie | NFluc of lactamase fused with dCas9, His tag allows for protein purification | Jiaofeng Li | |
| BBa_K1689014 | Coding | dCas9 fused with C terminal of beta-lactamase | Wei Weijia | |
| BBa_K1689015 | Coding | segment F[1,2] of DHFR-dCas9 fusion protein | Wei Weijia | |
| BBa_K1689016 | Coding | fusion protein of dCas9-F[1,2] | Wei Weijia | |
| BBa_K1689017 | Coding | DHFR F3 fragment-dCas9 fusion protein | Wei Weijia | |
| BBa_K1689018 | Coding | dCas9 fused with F3 | Wei Weijia | |
| BBa_K1689019 | Coding | Δα segment of β-galactosidase fused with dCas9 | Wei Weijia | |
| BBa_K1689020 | Coding | dCas9 fused with delta omega segment of β-galactosidase | Wei Weijia |
Parts for paired dCas9 reporter system
Nluc416-FRB (BBa_K1689003),
FKBP-Cluc398 (BBa_K1689005),
Nluc398-FRB (BBa_K1689004),
FKBP-Cluc394 (BBa_K1689006)
are used to validate the reconsitituation activity of split luciferase.
Nluc-dCas9(BBa_K1689010),
Cluc-dCas9(BBa_K1689009),
dCas9-Nluc (BBa_K1689008),
dCas9-Cluc (BBa_K1689007)
fusion proteins are used for constructing paired dCas9 reporter system.
Δα-dCas9 (BBa_K1689011),
dCas9-Nlact (BBa_K1689012),
Nlact-dCas9 (BBa_K1689013),
dCas9-Clact (BBa_K1689014),
F[1,2]-dCas9 (BBa_K1689015),
dCas9-F[1,2] (BBa_K1689016),
F3-dCas9 (BBa_K1689017),
dCas9-F3 (BBa_K1689018),
dCas9-Δα (BBa_K1689019),
Δω-dCas9 (BBa_K1689020)
are used for further development of dectection.
Parts for Molecular Beacon
STV-Nluc (BBa_K1689001),
Cluc-STV (BBa_K1689002)
are used for binding to molecular beacon.
Parts for sgRNA generation
sgRNA generator (BBa_K1689000) is used for generating sgRNA
Part Collection
2015 Peking iGEM has not only transformed the sequence information into detectable bioluminescence, but also explored a wide range in which Paired dCas9 (PC) Reporter System is able to be applied. Since we have chosen split enzyme as our reporter, it can be substituted by various kinds of enzymes, thus the form of read-out will be abundant at the same time. That is, our PC reporter system has successfully provided a platform to produce a variety of signals.
We combine the specific sequence binding activity of dCas9 with diverse characteristics of split enzymes, thus creating a part collection named "PC Reporters Collection".
The Collection includes:
dCas9-split luciferase Parts
Nluc-dCas9 (BBa_K1689010)
Cluc-dCas9 (BBa_K1689009)
dCas9-Nluc (BBa_K1689008)
dCas9-Cluc (BBa_K1689007)
dCas9-split Dihydrofolate Reductase Parts
F[1,2]-dCas9 (BBa_K1689015)
F3-dCas9 (BBa_K1689017)
dCas9-F[1,2] (BBa_K1689016)
dCas9-F3 (BBa_K1689018)
dCas9-split β–Lactamase Parts
dCas9-Nlact (BBa_K1689012)
Nlact-dCas9 (BBa_K1689013)
dCas9-Clact (BBa_K1689014)
dCas9-splitβ–Galactosidase Parts
Δα-dCas9 (BBa_K1689011)
dCas9-Δα (BBa_K1689019)
Δω-dCas9 (BBa_K1689020)
Among them, the PC reporters with split luciferase oxidizes luciferin and gives out bioluminescence signal, while the other three reporters take electric signal as their output, which is also able to be detected and qualified.