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...ling section was to create an integrated, kinetic model of our Cas9-guided chloroplast transformation mechanism. This model can be used to understand the internal <li>Cas9 cleavage on the chloroplast genome (for both on- and off-target sites)

...f time it takes (2-3 months) to achieve homoplasmy (when all the copies of chloroplast DNA are transformed by our cassettes and so they are stable and expressed a <center><p>On the other hand chloroplast engineering has a very bright future and it is definitely worth using. Chlo

...cks and building a library of modular genetic components for Chlamydomonas chloroplast.</p> ...mercial biolistic devices are very expensive. Finally, the lack of modular chloroplast genetic parts available makes gene cloning significantly more time-consumin

...h could help achieve essential homoplasmy (transformation of all copies of chloroplast DNA) in one generation instead of 2-3 months of selection. Our design incor ...50%; text-align:center;">Chlamydomonas, a single-celled alga with only one chloroplast, has its natural advantage over many other single-celled organisms in the l

...developed based on this microalga fully sequenced genomes (mitochondrial, chloroplast and nucleus). Other related advantages are its simple life-cycle and ease t ...me studies demonstrated also high heterologous protein accumulation in the chloroplast when using a psbA promoter and 5’UTR, only in psbA knockout strain (Manue

...150%; text-align:right">Due to extensive evolutionary conservation of the chloroplast genome, research in the chloroplasts of microalgae, such as Chlamydomonas r ...t-family:Open Sans; font-size:180%; text-align:center">III. Bottlenecks of chloroplast engineering

...ily:Arvo; font-size:110%">The Cambridge-JIC 2016 IGEM team has developed a chloroplast transformation toolkit to allow future teams to use plastids as a new chass ...tackle with our project. One of them is that homoplasmy, the last step of chloroplast transformation leading to the stable integration of the transgene in the g

...experts in the Plant Sciences Department and dived into the literature on chloroplast transformation. ...esised (primers surprisingly :D ) but we will try to amplify them from the chloroplast genome. Our plasmid design had at least 10 major changes of direction and w

...rest into as specific site in chloroplast DNA and then transform the other chloroplast DNA molecules with the same gene of interest in a way similar to transposon

...wetlab results revolve around the achievement of the first C. reinhardtii chloroplast library in phytobrick standard which, among others, allows to creation of a <p style="font-family:'Roboto Condensed'; font-size:150%">Firstly, chloroplast genomes have a high A/T content, therefore many could not be synthesized in

...s, Watford Grammar School for Girls, to deliver a talk about the future of chloroplast engineering to over 60 members of staff and science & maths students from y ...size:150%">This then led us onto the subject of our own project this year: chloroplast engineering in Chlamydomonas reinhardtii. We captivated the students’ ima

...promoter initiate the transcription of gene hhl1,so there will be HHL1 in chloroplast before PSⅡ is damaged，and the repair process can be faster.</div></p> ...div class="text_font">&nbsp;&nbsp;1）Under growth light condition, the chloroplast of the plants that transformed in four different vectors were not si

...or the generation of genetic circuits within the Chlamydomonas reinhardtii chloroplast. A chassis novel to the iGEM competition but which harnesses a massive pote ...nsed'; font-size:150%">In conclusion, we have proved our new Chlamydomonas chloroplast parts library which is functional and amplifiable in competent E. coli cell

...cells. In the future, we also aim to prove the gun’s use in carrying out chloroplast transformations, by attempting transformation of chlamydomonas samples. ...FP (green fluorescent protein), both codon-optimized for expression in the chloroplast.</p>

...ented on the registry, establishing the phytobrick standard in iGEM (first chloroplast part in the competition!). We hope this will kickstart the use of plastid s <li>Proved that our library of C. reinhardtii chloroplast L0 parts (first-ever in iGEM!) can be used to assemble higher-order constru

...to achieve as part of our project, through a standardised cas9 system for chloroplast engineering, which follows the phytobrick common syntax. Other relevant cha ...ct will help to further support our efforts in removing the bottlenecks to chloroplast engineering, by changing public reaction and accessibility to the technolog

...ity College London, for continuous advice in working towards a homoplasmic chloroplast transformation in Chlamydomonas .... Saul Purton</b>, University College London, for providing us a series of chloroplast transformation vectors and the TN72 Chlamydomonas reinhardtii strain, as we

...ts of 2 genes <i>psbT</i> and <i>psbB</i>. <i>psbT</i> encodes for a small chloroplast hydrophobic polypeptide associated with the core complex [14], while <i>psb <li>Takahashi, Y., et al., <i>Directed disruption of the Chlamydomonas chloroplast psbK gene destabilizes the photosystem II reaction center complex.</i> Plan

...etal in shape of cup, with or whitouth a pyrenoid in the part axial of the chloroplast with two or five granules of starch. <br>Their cells Wall is thin. Multipli

...p><div class="text_font">&nbsp;&nbsp;1）Under growth light condition, the chloroplast of the plants that transformed in four different vectors were not damaged, ...fferent level although the protein HHL1 exsist. So the red fluorescence of chloroplast had decreased, but the decrease level is quit different, the lower the fluo