sgRNA

Guide RNA interacting with target DNA. Image credit: William Reid

Guide-RNA Engineering – Enhancing Activity and Function

The functionality of Cas9 is determined in part by the guide RNA with which it is associated. Norwak et al.(2016) review briefly how guide RNAs can be modified to enhance their function and to functionalize them in novel ways. Norwak […]

Guide RNA interacting with target DNA. Image credit: William Reid

Intramolecular interactions in gRNA may influence Cas9 cleavage activity

In an article recently published in Nature Communications, Thyme et al (2016) discuss the interactions of the sequences of sgRNAs, the role of chromatin, and how these factors might interfere with Cas9 cleavage activity. By investigating the activity of Cas9 […]

Phosphorothioate oligomers increase knock-in frequency and precision in TALEN- and Cas9-mediated genome editing

In a recent paper by Renaud et al. (2016), the authors investigate methods to increase genome editing knock-in efficiency by modifying the chemistry of the donor oligomers to be resistant to nuclease activity. The modifications they tested included locked nucleic […]

Adult male spotted wing drosophila, Drosophilia suzukii (Matsumura). Photograph by Martin Hauser, California Department of Food and Agriculture.

Gene Editing in an Invasive Species

Li and Scott report their successful use of Cas9 to knockout two genes in Drosophila suzukii in a recent issue of Biochemical and Biophysical Research Communications. Drosophila suzukii or the spotted wing Drosophila although a native of Asia has recently […]

"Papilio machaon MHNT CUT 2013 3 11 Cahors Female" by Didier Descouens - Own work. Licensed under CC BY-SA 4.0 via Commons - https://commons.wikimedia.org/wiki/File:Papilio_machaon_MHNT_CUT_2013_3_11_Cahors_Female.jpg#/media/File:Papilio_machaon_MHNT_CUT_2013_3_11_Cahors_Female.jpg

CRISPR/Cas9 Gene Editing in Papilionid Butterflies

Li et al (2015) recently published an exciting paper on butterflies genomics and functional genomics . Li et al have worked towards producing high quality genomes for extremely heterozygous and closely related butterflies P. xuthus and P. machaon. Further, they […]

Cas9 + sgRNA

CRISPRscan – A Tool Aiding Selection of CRISPR-Cas9 gRNAs

Just published online in Nature Methods, Moreno-Mateos et al report testing the activities of >1200 gRNAs in zebrafish embryos and used the results to create an algorithm for predicting gRNA efficacy. The advent of the CRISPR-Cas9 system has revolutionized genome […]

Cas9 with functional domains highlighted in relation to target DNA and guideRNA

CRISPR-STAT to Evaluate Cas9/guideRNAs

Carrington et al (2015) describe another approach to functionally assessing guide RNAs for use with Cas9  that is quick, relatively easy and inexpensive. After designing and assembling guide RNAs to be used to direct Cas9 to specific targets, investigators would […]

Structure of a TALE binding to DNA

Optimized TALEN Application and Screening in Drosophila melanogaster

Lee et al 2015 provide a detailed methodology for gene editing using TALENs in Drosophila melanogaster. While TALEN use is relatively well established in D. melanogaster, fine tuning the injection concentrations of designer nucleases allows for the maximum efficiency in […]

Electroporation Theory

Electroporating the CRISPR/Cas9 System

Qin et al. (2015) describe their successful efforts to use electroporation technology to introduce CRISPR/Cas9 system components into mouse embryos and while not exactly insect biology their motivations for pursuing this approach are shared by many insect biologists with an interest […]

CRISPR/Cas9 Knock-in Technology

Cas9 Against RNA Targets In Vivo

Price et al (2015) have recently published a paper in PNAS in which they report that Cas9 can be used as a RNA-targeting system. Clustered, regularly interspersed, short palindromic repeats-CRISPR associated (CRISPR-Cas) systems are a diverse collection of adaptive immune […]

Delivering gene-editing proteins in vivo using a new strategy.

Delivering genetic technologies to appropriate cells at the appropriate times is, for many insect systems, the biggest obstacle to the widespread use of these technologies. Microinjecting developing embryos with DNA, RNA or protein remains the dominant mode of delivery in […]

“CHOPCHOP”ing with CRISPRs and TALENs: A New Web Resource for Genome Editing Design

With the growing influence and applicability of genome editing technologies, TALENs and more recently CRISPRs, offer genetic capabilities to an ever growing list of organisms. In turn, as more and more researchers become interested in using these technologies, a common […]