Traditional genome editing in insects involves directly injecting embryos. However, most insect embryos are small and sensitive to manipulation.
Cas9
High Fidelity SaCas9
CRISPR-Cas9 proteins have widely been used for a variety of genome editing applications, albeit with serious off-target activities.
Efficient Knock-ins With Short Homology Arms
In a recent publication in eLIFE, Kanca et al (2019) describe three methods to create donors for efficient CRISPR-based insertion of homology donors in vitro and in vivo.
Gene Editing Screwworms and Blowflies
Paolo et al. report the first successful use of CRISPR/Cas9 in the New World Screwworm fly, Cochliomyia hominivorax (C. hominivorax), a parasite of warm blooded animals including humans, as well as a CRISPR/Cas9 protocol for the Australian sheep blowfly, Lucilia […]
Homology-Free Knockins in Insects
Bosch et. al. 2019 report in biorxiv a homology-independent genomic site-specific gene knock-in method that can be applied both in vitro and in vivo. Traditional methods of site-specific knock-in use homologous repair, which requires cloning of long homology arms that […]
Small Nucleic Acid Based Inhibitors of SpCas9 (SNuBs)
Barkau et. al. 2019 report the development of small nucleic acid-based inhibitors (SNuBs) of SpCas9. Due to considerations such as off-target activity, it is desirable that we have a high degree of control over CRISPR/Cas9 cleavage rates. However, while several […]
CRISPR/Cas9-Mediated NHEJ Mutations in Anastrepha suspensa Paves Way for HDR
In an a recent experiment published in Gene, Li and Handler (2019) utilized readily available study systems and newly emerging genetic technologies and demonstrated both germline and somatic CRISPR/Cas9 induced mutations in Anastrepha ludens. The creation of heritable mutations was […]
Heritable Gene Editing in Thermobia domestica
Ohde et al report the first successful development and use of CRISPR/Cas9 in the basally branching insect Thermobia domestica. By co-injecting Cas9 nuclease with sgRNA targeting a T. domestica ortholog of the Drosophila melanogaster white gene (Td-w), the authors created […]
Microinjection of the Sand Fly Lutzomyia longipalpis, a Vector for Leishmaniasis
Two recent papers, Jefferies et al (2018) and Martin-Martin et. al (2018), describe experiments to develop and optimise a sand fly microinjection protocol. There are 2 authors to this post: Alla Madich, Simon Collier, technical constraints on this website allow […]
Cas9 Orthologs with Minimal PAM Requirements
In their recent paper Chatterjee and colleagues (2018) describe a natural protospacer adjacent motif (PAM) plasticity of previously uncharacterized Cas9 endonuclease from Streptococcus canis (ScCas9). Chatterjee et al (2018) report ScCas9’s affinity to minimal 5’-NNG-3’ PAM sequences and describe its accurate editing capabilities, […]