CRISPR

standard Call for Sweeping Moratorium on Gene Drive Research and Application at UN Biodiversity Conference

The UN Convention on Biological Diversity is currently holding their Biodiversity Conference in Cancun, Mexico.  Monday 160 organizations from six continents released a statement calling for a global moratorium on all work on gene drives, including laboratory research.   The […]

Male Locusta migratoria migratorioides photographed in Katavi National Park, Tanzania

standard Orco Knockout Mutants of Locusta migratoria

‎  A recent report by Li et al. demonstrates that by using a state-of-the-art CRISPR/Cas9 genome editing, it is now possible to study the function of Orco in the locust, Locusta migratoria .  The gene odorant receptor co-receptor (Orco) encodes […]

standard Genetically Modified Plant-Feeding Insects: A DARPA Project

When transgenic mosquito technology was being developed an idea was floated to use genetically modified mosquitoes as flying syringes to deliver important antigens to immunize people against various pathogens.  Informed consent among other things made this an overly complex problem […]

standard Dominant Maternal Effects Caused by CRISPR/Cas9

Expressing Cas9 and guide RNAs from transgenes integrated into the genome of organisms typically leads to higher rates of mutagenesis of the target gene compared to when those components are provided transiently by microinjection, for example. Lin and Potter describe […]

standard Loss of Function Mutations – Approaches, Limitations

Technologies are democratizing insect genetics and it seems as if everyone is thinking about making ‘knockouts’.  This is a good thing but as Housden et al. nicely explain in a recent paper in Nature Reveiws Genetics explain: “although LOF (loss […]

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

standard 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 […]

standard CRISPR-SD, a programmable Cas9 approach to generating X-shredder lines in insects with XY mating systems

In a recent paper in Scientific Reports, Galizi, Hammond, et al report on the use of CRISPR-Cas9 to develop a sex-distortion system in Anopheles gambiae, which they termed CRISPRSD. Earlier, the group had demonstrated that transgenic An. gambaie expressing the […]

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standard Gene Editing Spodoptera

Three papers recently described successful CRISPR/Cas9 mutagenesis in two species of Spodoptera.  Spodoptera litura and S. littoralis. Bi et al (2016) use Cas9 to create somatic mutations in S. litura in developmental gene abdominal-A, abd-A, and analyzed the resulting larvae […]

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

standard 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 […]

A selfish gene (blue) spreading through a mosquito population

standard U.S. National Academy of Science’s Report on Gene Drive Research

The United States National Academy of Sciences published a report entitled Gene Drives on the Horizon: Advancing Science, Navigating Uncertainty, and Aligning Research with Public Values recently.  The report was written by the Committee on Gene Drive Research in Non-Human […]

standard 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 […]

A selfish gene (blue) spreading through a mosquito population

standard Insect Gene Drives – Reviewed and Compared

Champer et al have published a review on the topic of engineered gene drives to manipulate wild populations in a recent Nature Reviews Genetics. Recent attention on Cas9-based gene drive systems has raised the awareness of these types of genetic […]

Danaus plexippus By William Warby (https://flic.kr/p/dtKZF2) [CC BY 2.0 (http://creativecommons.org/licenses/by/2.0)], via Wikimedia Commons

standard Monarch Reverse Genetics

Markert et al. just reported their effective use Cas9 and TALENs to knock out cryptochrome 2 and clock in Danaus plexippus using some strategies and approaches that are generally useful to insect scientists and much more efficient than earlier reports […]

standard Animal and Insect Transgenics – Parallels

James Murray and Elizabeth Maga recently recounted briefly the development and evolution of animal transgenic technologies in a recent issue of Transgenic Research and the similarities and differences between the development and evolution of insect transgenic technologies, which were ongoing […]

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

standard 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 […]

standard Enhancing Homology Repair-Mediated Genome Editing

In a recent paper in Nature Biotechnology, Richardson et al (2016) report on the interaction of target-site bound Cas9 and dCas9 on DNA strand accessibility- and have identified an optimized approach to increase homologous recombination (HR) mediated repair. They found […]

standard Another High-Fidelity SpCas9 Nuclease

Kleinstiver et al. describe in Nature their successful efforts to create high-fidelity SpCas9 nuclease variants that have high on-target efficiencies and low or no off-target effects.  The results are SpCas9-HF1. When using Cas9 or other engineered nuclease systems one hopes […]

Synthetic gene drive system based on site specific DNA endonucleases (homing endonucleases, ZFN, TALENS, Cas9). Target sites and homology arms are chosen so that the endonuclease and associated genes (gRNA and other transgenes) are copied into homologous chromosomes.

standard An Overview of Gene Drive

Gantz and Bier reflect on and consider the implications of Cas9-based genome editing on genetics in a recent Bioessays that is worth reading if you are trying to keep up on this rapidly evolving part of the genetics ecosystem. Not […]

guide RNA relative to target sequence

standard Homology independent CRISPR Knock-ins

Lackner et al., in an effort to eliminate the need to synthesize multiple, unique homology based templates for Cas9-mediated homologous recombination, report the use of a universal donor template that will insert tags into potentially any genomic site in the human […]

Parhyale hawaiensis

standard Somatic Editing in Parhyale hawaiensis for Loss-of-Function Analysis

In a recent paper in Current Biology, IGTRCN Participant Arnaud Martin and colleagues use Cas9 gene editing technology to study the development of the crustacean Parhyale hawaiensis.  In this study by Martin et al., the specific morphology of serially homologous […]

Anopheles gambiae

standard Gene Drive In Anopheles gambiae

Hammond et al.’s just published paper in  Nature Biotechnology is the second paper in a month demonstrating effective CRISPR-based gene drive in a mosquito vector of malaria.  In this case the main malaria vector in Africa, Anopheles gambiae. Gene drive […]

standard Cas9-Mediated Gene-Drive In Mosquitoes

Gantz and Bier who demonstrated the fundamental principles of a Cas9-based gene drive system using Drosophila melanogaster in a publication earlier this year have recreated that system in the mosquito Anopheles stephensi with the help of collaborators.  Gantz et al. […]

standard Dopamine receptor knock-outs in crickets

Awata et al. (2015) have used Cas9 to knock-out Dop1 in Gryllus bimaculatus to assess the role of dopamine neurons in mediating reward and punishment. Functional genomics technologies like transposons and first and second generation gene-editing technologies have been available […]

Harmonia axyridis, 19-spot form

standard Sawfly and Ladybird Beetle Gene Editing

Hatakeyama et al (2015) have successfully used gene editing technology in Athalia rosae (sawfly) and Harmonia axyridis (ladybird beetle), enlarging the range of insects in which these technologies have been applied.  These results are reported in Insect Molecular Biology. Hatakeyama […]

standard Compendium of Genome Engineering

Koen Venken and colleagues have just published an excellent review of ‘Genome Engineering: Drosophila melanogaster and beyond‘ that is comprehensive and highly informative, and should serve as a useful reference for anyone interested in genome modification techniques with nucleotide precision […]

"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

standard 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

standard 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 […]

standard Editing Tribolium’s Genome

Gilles et al (2015) recently published a paper in Development describing CRISPR-mediated gene targeting and gene replacement in the flour beetle Tribolium castaneum. The authors started with the Tribolium line Pig-19, which strongly expresses enhanced green fluorescent protein (EGFP) in […]

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

standard 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 […]

Basics of Cas9 Activity

standard Cas9 Gene Drive & Classical Biological Control

Webber et al have published an Opinion essay in PNAS concerning the growing interest in using engineered endonucleases (Cas9 in particular) to create gene drive systems to combat invasive species. Homing endonucleases are well known and well studied site-specific endonucleases […]

Structure of a TALE binding to DNA

standard 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 […]

standard Microinjection independent genome engineering

Takahashi et al. (2105) describe a microinjection independent genome engineering method for mice that is an interesting innovation that seeks to address technical challenges associated with the creation of transgenic and genetically modified mice that have good analogs in insect […]

standard Powerful Genetic and Genomic Toolbox of Nasonia Wasps

Jeremy Lynch (2015) has published a nice overview of the genetic/genomic tools currently available for Nasonia species. Insects in the order of Hymenoptera are emerging as new genetic and genomic tools for evolutionary biology, development, and behavioral ecology. Among over […]

Electroporation Theory

standard 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 […]

standard CRISPR with Independent Transgenes or Mutagenic Chain Reaction for Creating Homozygotes in Lab?

In fall of 2014, two publications came from George Church’s group (Esvelt et al., 2014; Oye et al., 2014) postulating on the potential for the CRISPR/Cas9 system to be used as an autonomous gene drive for various pest management goals. […]

standard Short Course on Insect Genetic Technologies – August 2015

The Insect Genetic Technologies Research Coordination Network is offering a short course on insect genetic technologies August 17-21, 2015. Today there are myriad technologies for modifying genomes. Technologies are emerging rapidly and it is a challenge for investigators to keep […]

CRISPR/Cas9 Knock-in Technology

standard 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 […]

standard CRISPR-Cas9 mediated “Mutagenic Chain Reaction”

A recent paper in Science by Gantz and Bier (2015) investigated the potential of utilizing the CRISPR-Cas9 system as, essentially, a programmable homing endonuclease to generate homozygous loss-of-function (LOF) mutants. Advances in genome editing have allowed for the directed generation of […]

standard Exchangeable Cassettes Controlled by Endogenous Genes

Two recent publications, Gnerer et al. and Diao et al., address the issue of recreating the fine transcriptional patterns of endogenous genes with binary expression systems in insects. Binary expression systems are extremely powerful tools for controlled gene manipulation. The […]

standard Pulling Recent CRISPR/Cas9 Work in Drosophila Together

Xu et al. review the recent history of using CRISPR/Cas9-based technologies in Drosophila melanogaster. Within the last two years there have been numerous publications describing the use of CRISPR/Cas9-based technologies in Drosophila melanogaster.  These last two years have been a time […]

FlyFacility, Cambridge University. A insect genetic technologies service provider and resource

standard The Fly Facility at the University of Cambridge Department of Genetics

The Department of Genetics at the University of Cambridge is primarily located in a large Edwardian building close to the centre of town. The department has a long and distinguished history of Drosophila research beginning with John Thoday’s evolutionary genetics […]

standard The Drosophila RNAi Screening Center (DRSC) – A functional genomics resource useful to all insect biologists.

The Drosophila RNAi Screening Center (DRSC) at Harvard Medical School was founded by Prof. Norbert Perrimon in 2003 to provide community support for genome-wide RNAi screens in cultured Drosophila cells. Now, more than a decade later, we are still going […]

Cas9 + sgRNA

standard CRISPR-Cas9 Through Eyes of Two Leaders in the Field

In a comprehensive and definitive review published in Science, Jennifer Doudna and Emmanuelle Charpentier, two scientist who made seminal discoveries leading to the current boom in genome engineering, walk us through the (brief) history, scientific applications, and future promise of […]

standard Comparing TALE- and dCas9-Based Designer Transcription Factors

Genome editing with programmable nucleases using transcription activator-like effector nucleases (TALENs) or RNA-guided clustered regularly interspersed short palindromic repeat (CRISPR) associated protein (Cas9) have become popular and important technologies. These systems can be modified to carry other functional protein domains […]

standard IGTRCN Trainees – Fall 2014!

The IGTRCN is working to diffuse knowledge and technology from those who ‘know’ and ‘have’ to those who do not but need to. The Peer-to-Peer Training Fellowships are intended to facilitate the process by providing modest resources to encourage interactions. […]

standard 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 […]

standard Plasmids for Rapid and Versatile Genome Editing

As is the case for many other organisms, CRISPR has rapidly become the method of choice for targeted genome modification in insects. Much of the method development has been taking place in the model organism Drosophila melanogaster, where, in little […]

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

standard CRISPR/Cas9 for RNA Cleavage

The CRISPR/Cas9 system is quickly becoming the most versatile method for modifying DNA targets, but researchers seeking equivalent control against RNA transcripts have been left behind. In a new paper by O’Connell et. al, (2014) the authors describe a clever […]

standard TGV: Programmable DNA Binding Proteins for Live Visualization of Chromatin

Transcription activator-like effector (TALE) technologies are being used in a variety of creative ways by swapping functional protein domains that are connected to the ‘programmed’ DNA binding domain.  Miyanari (2014) and Miyanari et al (2013) describe their use of TALEs […]

standard A Collection of Methods for Editing Insect Genomes

The August 2014 issue of METHODS is devoted to the topic of “Editing and Investigating Genomes with TALE and CRISPER/Cas Systems: Genome Engineering Across Species Using TALENS” Five of the 14 protocols included in this issue are specifically focused on […]

standard Transgenic Underdominance: Pushing Transgenes into Populations.

Insect population replacement and suppression using transgenic strategies are being considered for insects of medical and agricultural importance. Rapidly increasing the frequency of transgenes in target populations after transgene release using genetic drive systems remains a significant challenge. Driving transgenes […]

standard “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 […]

Structure of a TALE binding to DNA

standard TAL Effector Modulators: Beyond Nucleases

Programmable DNA-binding proteins such as Zinc Finger Nucleases, TALEs and CRISPR/Cas9 are versatile tools for genome modification. While mutagenesis (knock outs and knock ins) has been a major initial objective in using these systems to date, it is clear that […]

standard Synthetic Selfish DNA as Genetic Technologies

  Selfish genes are genes that can spread themselves rapidly within genomes and populations.   Think – transposable elements, B chromosomes, homing endonucleases, selfish sex chromosomes, genomic exclusion to name a few. Some are promising candidates for the genetic engineering […]

Cas9 + sgRNA

standard Purified Cas9 Protein for Insect Genome Modification with CRISPRs

The interest in CRISPR/Cas9 genome-editing and -modification technologies is quite extensive and there has been a flood of data dealing with optimizing efficiency among other things.               In insect systems there are a number […]

standard Insect Genetic Technologies Service Providers

Insect Genetic Technology Service Providers If you would like to add to this list use the “Make a Submission” button and select “Knowledgebase” For insects other than Drosophila melanogaster Non-Commercial The Institute for Bioscience and Biotechnology Insect Transformation Facility, University […]

Meiotic drive in Drosophila melanogaster

standard The Road Ahead for Using Gene Drive Systems

Emerging technologies have employed chemical, ecological and genetic strategies for effective and economical pest management. The various strategies around the transgenic augmentation of pests has shown great promise in being able to combat vector borne diseases and eradicate agricultural pests […]

XOR Circuit: You get output only when there is input from A or B but not both or neither.

standard CRISPy Circuits

Sophisticated tools for genome modification allow investigators to devise ever-increasing complex genetic circuits. The Gal4 /UAS system is one example of a simple binary system. While powerful, this system does not allow for particularly complex genetic circuits to be assembled. […]

TALENs and CRISPRs

standard Multigene Mutagenesis of Silk Moth with CRISPRs.

Genome editing using CRISPR/Cas9 is changing the way we think about genome modification. The CRISPR system, because it relies on a small user-provide guide RNA (gRNA) to give the Cas9 endonuclease its sequence specificity, is easy to use, relative to, […]

vasa Cas9 expressing Drosophila. From Xue et al. 2014

standard Vasa Cas9 Experession Yields Good Mutation Rates in Drosophila

Xue and colleagues report efficient knock-out and knock-in manipulations of the Drosophila genome with a Cas9 transgenic system. Others have made similar observations.  Xue et al. report the germline mutational rate ranging from 14% to 95%. They also demonstrated the […]

Uses for nuclease defective CRISPRS

standard CRISPR/Cas9 in Drosophila – Other Insects not far behind.

While genome editing technologies have been available for the past decade or so, until recently they have been challenging and expensive to use. The Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR/Cas9) system is changing all of that. This bacterial defense […]