↓ Skip to main content

A redesigned CRISPR/Cas9 system for marker-free genome editing in Plasmodium falciparum

Overview of attention for article published in Parasites & Vectors, April 2016
Altmetric Badge

About this Attention Score

  • Above-average Attention Score compared to outputs of the same age (62nd percentile)

Mentioned by

twitter
4 tweeters

Citations

dimensions_citation
28 Dimensions

Readers on

mendeley
129 Mendeley
You are seeing a free-to-access but limited selection of the activity Altmetric has collected about this research output. Click here to find out more.
Title
A redesigned CRISPR/Cas9 system for marker-free genome editing in Plasmodium falciparum
Published in
Parasites & Vectors, April 2016
DOI 10.1186/s13071-016-1487-4
Pubmed ID
Authors

Junnan Lu, Ying Tong, Jiaqiang Pan, Yijun Yang, Quan Liu, Xuefang Tan, Siting Zhao, Li Qin, Xiaoping Chen

Abstract

A highly efficient CRISPR/Cas9-based marker-free genome editing system has been established in Plasmodium falciparum (Pf). However, with the current methods, two drug-selectable markers are needed for episome retention, which may present hurdles for consecutive genome manipulations due to the limited number of available selectable markers. The loading capacity of donor DNA is also unsatisfactory due to the large size of the Cas9 nuclease and sgRNA co-expression system, which limits the size of knock-in DNA fragments. Because of the inefficient end joining (EJ) DNA repair mechanism of Pf, a suicide-rescue approach could be used to address the challenges. Cas9 nuclease and sgRNA were co-expressed from a single plasmid (suicide vector) with one selectable marker, and the donor DNA was ligated into the other plasmid (rescue vector) containing only the ampicillin-resistance gene (AmpR) and a ColEl replication origin (ori). Nonetheless, whether this approach can mediate even the regular gene editing in Pf remains unknown. This study aimed to demonstrate the basic gene editing function of this Cas9-mediated suicide-rescue system. The suicide and rescue vectors were constructed and co-transfected into Pf3D7. This system worked as expected when used to disrupt the Pfset2 gene and to insert a green fluorescent protein-renilla luciferase (gfp-ruc) fusion gene cassette of 3334 base pairs (bp) into the Pf47 locus, demonstrating that the suicide vector actually induced double-strand breaks (DSBs) and that the rescue vector functioned without maintenance via drug selection. The adapted marker-free CRISPR/Cas9 system with only a single episome-selectable marker performs well as the current systems for general gene editing which lays a solid foundation for further studies including consecutive gene manipulations and large gene knock-ins.

Twitter Demographics

The data shown below were collected from the profiles of 4 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 129 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 129 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 46 36%
Student > Master 19 15%
Researcher 18 14%
Student > Bachelor 15 12%
Student > Doctoral Student 6 5%
Other 13 10%
Unknown 12 9%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 56 43%
Agricultural and Biological Sciences 40 31%
Immunology and Microbiology 9 7%
Medicine and Dentistry 3 2%
Pharmacology, Toxicology and Pharmaceutical Science 2 2%
Other 3 2%
Unknown 16 12%

Attention Score in Context

This research output has an Altmetric Attention Score of 3. 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 17 April 2016.
All research outputs
#7,381,315
of 14,448,829 outputs
Outputs from Parasites & Vectors
#1,322
of 3,897 outputs
Outputs of similar age
#99,010
of 264,092 outputs
Outputs of similar age from Parasites & Vectors
#1
of 1 outputs
Altmetric has tracked 14,448,829 research outputs across all sources so far. This one is in the 48th percentile – i.e., 48% of other outputs scored the same or lower than it.
So far Altmetric has tracked 3,897 research outputs from this source. They receive a mean Attention Score of 4.6. This one has gotten more attention than average, scoring higher than 65% 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 264,092 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 62% 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