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Seamless editing of the chloroplast genome in plants

Overview of attention for article published in BMC Plant Biology, July 2016
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  • In the top 25% of all research outputs scored by Altmetric
  • Good Attention Score compared to outputs of the same age (77th percentile)

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10 tweeters

Citations

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16 Dimensions

Readers on

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73 Mendeley
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Title
Seamless editing of the chloroplast genome in plants
Published in
BMC Plant Biology, July 2016
DOI 10.1186/s12870-016-0857-6
Pubmed ID
Authors

Elena Martin Avila, Martin F. Gisby, Anil Day

Abstract

Gene editing technologies enable the precise insertion of favourable mutations and performance enhancing trait genes into chromosomes whilst excluding all excess DNA from modified genomes. The technology gives rise to a new class of biotech crops which is likely to have widespread applications in agriculture. Despite progress in the nucleus, the seamless insertions of point mutations and non-selectable foreign genes into the organelle genomes of crops have not been described. The chloroplast genome is an attractive target to improve photosynthesis and crop performance. Current chloroplast genome engineering technologies for introducing point mutations into native chloroplast genes leave DNA scars, such as the target sites for recombination enzymes. Seamless editing methods to modify chloroplast genes need to address reversal of site-directed point mutations by template mediated repair with the vast excess of wild type chloroplast genomes that are present early in the transformation process. Using tobacco, we developed an efficient two-step method to edit a chloroplast gene by replacing the wild type sequence with a transient intermediate. This was resolved to the final edited gene by recombination between imperfect direct repeats. Six out of 11 transplastomic plants isolated contained the desired intermediate and at the second step this was resolved to the edited chloroplast gene in five of six plants tested. Maintenance of a single base deletion mutation in an imperfect direct repeat of the native chloroplast rbcL gene showed the limited influence of biased repair back to the wild type sequence. The deletion caused a frameshift, which replaced the five C-terminal amino acids of the Rubisco large subunit with 16 alternative residues resulting in a ~30-fold reduction in its accumulation. We monitored the process in vivo by engineering an overlapping gusA gene downstream of the edited rbcL gene. Translational coupling between the overlapping rbcL and gusA genes resulted in relatively high GUS accumulation (~0.5 % of leaf protein). Editing chloroplast genomes using transient imperfect direct repeats provides an efficient method for introducing point mutations into chloroplast genes. Moreover, we describe the first synthetic operon allowing expression of a downstream overlapping gene by translational coupling in chloroplasts. Overlapping genes provide a new mechanism for co-ordinating the translation of foreign proteins in chloroplasts.

Twitter Demographics

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Mendeley readers

The data shown below were compiled from readership statistics for 73 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
New Zealand 1 1%
United States 1 1%
Netherlands 1 1%
Unknown 70 96%

Demographic breakdown

Readers by professional status Count As %
Researcher 21 29%
Student > Ph. D. Student 14 19%
Student > Bachelor 10 14%
Student > Master 6 8%
Other 3 4%
Other 7 10%
Unknown 12 16%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 27 37%
Agricultural and Biological Sciences 27 37%
Business, Management and Accounting 1 1%
Environmental Science 1 1%
Unknown 17 23%

Attention Score in Context

This research output has an Altmetric Attention Score of 7. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 10 January 2019.
All research outputs
#2,486,601
of 14,122,258 outputs
Outputs from BMC Plant Biology
#202
of 2,009 outputs
Outputs of similar age
#59,858
of 264,479 outputs
Outputs of similar age from BMC Plant Biology
#1
of 1 outputs
Altmetric has tracked 14,122,258 research outputs across all sources so far. Compared to these this one has done well and is in the 82nd percentile: it's in the top 25% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 2,009 research outputs from this source. They receive a mean Attention Score of 3.1. This one has done well, scoring higher than 89% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 264,479 tracked outputs that were published within six weeks on either side of this one in any source. This one has done well, scoring higher than 77% of its contemporaries.
We're also able to compare this research output to 1 others from the same source and published within six weeks on either side of this one. This one has scored higher than all of them