Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant &lt;em&gt;Harpegnathos saltator &lt;/em&gt;

Sieber, Kayli; Saar, Maya; Opachaloemphan, Comzit; Gallitto, Matthew; Yang, Huan; Yan, Hua

doi:10.3791/61930

Cited by 7 publications

(8 citation statements)

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“…In their experiment, about 60% of the uninjected eggs hatched on petri dishes (our hatching rate was 22% with nursing workers) and about 50% of the larvae survived with adult nursing workers (none of our uninjected larvae matured to adulthood, while one water injected (0.5% of the eggs) and 15 CRISPR-injected (1.2% of the eggs) matured) Yan et al (2017). andSieber et al (2021) removed the eggs from the tape after injection, washed them with ethanol and incubated them on agar plate treated…”

mentioning

confidence: 84%

“…When compared to the four other CRISPR experiments conducted on ants (Chiu et al, 2020;Sieber et al, 2021;Trible et al, 2017;Yan et al, 2017), there are pros and cons to our approach.…”

Section: Comparison To Other Crispr Experiments Conducted On Ants And...mentioning

confidence: 99%

“…The first insect was the fruit fly Drosophila melanogaster (Gratz et al, 2013 ) and after that, for example, the silk moth Bombyx mori and the honeybee Apis mellifera (Cui et al, 2017 ). CRISPR‐Cas9 has been applied in at least three ant species prior to this study: twice in Harpegnathos saltator (Sieber et al, 2021 ; Yan et al, 2017 ), once in Ooceraea biroi (Trible et al, 2017 ) and once in Solenopsis invicta (Chiu et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

“…Eggs and larvae require workers' attention and for example, individuals with modified olfactory receptors had lower fitness in the colony (Trible et al, 2017 ). In these studies, different methods were developed to rear larvae into adults (Sieber et al, 2021 ; Trible et al, 2017 ; Yan et al, 2017 ) or the success of the gene modification was detected by sequencing at the larval stage (Chiu et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

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Genetic modification of the ant Lasius niger using CRISPR‐Cas9 technology

Konu

Kulmuni

Viljakainen

2022

Insect Molecular Biology

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CRISPR-Cas9 has become one of the most prominent gene editing tools available and it has been utilized in various organisms from bacteria to fungi, plants, and animals. In this study, we developed a CRISPR-Cas9 protocol for the black garden ant Lasius niger, a common and easily available study species for lab and field experiments. To create indel mutations using CRISPR-Cas9 in L. niger, we targeted three different locations in a wellstudied eye pigmentation gene cinnabar, generating several mutations that disrupt the ommochrome biosynthesis pathway and result in the lack of the pigment and therefore, abnormal eye coloration in adult workers. We also developed a protocol to collect L. niger eggs, inject them with CRISPR-Cas9 construct, and rear the eggs into mature adult workers with the assistance of nursing workers. We demonstrated for the first time in L. niger that CRISPR-Cas9 is an excellent tool to create targeted mutations for this species. Our protocol can be referred to when developing similar studies for other species of ants and eusocial insects.

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confidence: 84%

“…When compared to the four other CRISPR experiments conducted on ants (Chiu et al, 2020;Sieber et al, 2021;Trible et al, 2017;Yan et al, 2017), there are pros and cons to our approach.…”

Section: Comparison To Other Crispr Experiments Conducted On Ants And...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Genetic modification of the ant Lasius niger using CRISPR‐Cas9 technology

Konu

Kulmuni

Viljakainen

2022

Insect Molecular Biology

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“…Establishment of genetic tools in eusocial insects continues to expand into new species. The CRISPR/Cas9 system has notably been established in three ant species, H. saltator [167,168], O. biroi [169], and most recently in the fire ant S. invicta [170]. CRISPR/Cas9 [171][172][173][174][175][176] and transgenesis [177,178] have also been established in the honeybee A. mellifera.…”

Section: Eusocial Insects As Modelsmentioning

confidence: 99%

(Epi)Genetic Mechanisms Underlying the Evolutionary Success of Eusocial Insects

Sieber

Dorman

Newell

et al. 2021

Insects

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Eusocial insects, such as bees, ants, and wasps of the Hymenoptera and termites of the Blattodea, are able to generate remarkable diversity in morphology and behavior despite being genetically uniform within a colony. Most eusocial insect species display caste structures in which reproductive ability is possessed by a single or a few queens while all other colony members act as workers. However, in some species, caste structure is somewhat plastic, and individuals may switch from one caste or behavioral phenotype to another in response to certain environmental cues. As different castes normally share a common genetic background, it is believed that much of this observed within-colony diversity results from transcriptional differences between individuals. This suggests that epigenetic mechanisms, featured by modified gene expression without changing genes themselves, may play an important role in eusocial insects. Indeed, epigenetic mechanisms such as DNA methylation, histone modifications and non-coding RNAs, have been shown to influence eusocial insects in multiple aspects, along with typical genetic regulation. This review summarizes the most recent findings regarding such mechanisms and their diverse roles in eusocial insects.

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