https://www.ncbi.nlm.nih.gov/pubmed/27715425?dopt=Abstract&utm_source=dlvr.it&utm_medium=twitter

*Abstract*

The European honeybee (Apis mellifera L.) is used as a model organism in
studies of the molecular and neural mechanisms underlying social behaviors
and/or advanced brain functions. The entire honeybee genome has been
sequenced, which has further advanced molecular biologic studies of the
honeybee. Functions of genes of interest, however, remain largely to be
elucidated in the honeybee due to the lack of effective reverse genetic
methods. Moreover, genetically modified honeybees must be maintained under
restricted laboratory conditions due to legal restrictions, further
complicating the application of reverse genetics to this species. Here we
applied CRISPR/Cas9 to the honeybee to develop an effective reverse genetic
method. We targeted major royal jelly protein 1 (mrjp1) for genome editing,
because this gene is predominantly expressed in adult workers and its
mutation is not expected to affect normal development. By injecting sgRNA
and Cas9 mRNA into 57 fertilized embryos collected within 3 h after
oviposition, we successfully created six queens, one of which produced
genome-edited male offspring. Of the 161 males produced, genotyping
demonstrated that the genome was edited in 20 males. All of the processes
necessary for producing these genome-edited queens and males were performed
in the laboratory. Therefore, we developed essential techniques to create
knockout honeybees by CRISPR/Cas9. Our findings also suggested that mrjp1
is dispensable for normal male development, at least till the pupal stage.
This new technology could pave the way for future functional analyses of
candidate genes involved in honeybee social behaviors.

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