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**This page is a work-in-progress concept of reformatting our resource and reagent list.**
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Full Table of Contents
CRISPR
gRNA Design Resources
Resource | Description | Author/Source |
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CRISPR Optimal Target Finder | Identifies gRNA targets within a provided sequence and additionally searches genome-wide (release 6, current FlyBase release) for potential off-target sites. Includes numerous Drosophila species. Incorporates PAM sequence detection. | O'Connor-Giles, Wildonger, and Harrison Labs University of Wisconsin-Madison WI, USA |
CRISPRscan | Searchable and browsable collection of genome-wide (release 6, current FlyBase release) sgRNA target sites. Displays sgRNA sites as tracks on UCSC's genome browser and allows searching via gene or prediction via sequence. Support for numerous species (fly, zebrafish, mouse, human, others). Incorporates PAM sequence detection. | Giraldez Lab Yale University New Haven, CT, USA |
DRSC Find CRISPR Tool | A tool for discovering gRNA targets located throughout the fly genome (release 5) using gene IDs, symbols, or chromosome locations. Allows for filtering of off-target sites and control of mis-match stringency. Incorporates PAM sequence detection. | DRSC Harvard Medical School Boston, MA, USA |
E-CRISPR | gRNA genome-wide (release 5) target finder, searchable via gene symbol or sequence. Support for numerous species (fly, zebrafish, mouse, human, others). Offers "relaxed, medium and strict" search options. Incorporates PAM sequence detection. | DKFZ/Boutros Lab Heildelberg, Germany |
CHOPCHOP | Search for gRNA targets (release 5). Flexible inputs (gene name, genomic coordinates or DNA sequence) and detection of off-target sites. Incorporates PAM sequence detection. | Schier & Church Labs Harvard University Cambridge, MA, USA |
CRISPR Design | Identification of gRNA sites genome wide (release 5). Offers options for multiple species. Incorporates PAM sequence detection. | Zhang Lab MIT Boston, MA, USA |
CRISPOR | A program that helps design, evaluate, and clone guide sequences for the CRISPR/Cas9 system. Allows for searches across multiple species of Drosophila, including melanogaster (release 5). | Tefor France |
CRISPR Stocks
Resource | Description | Author/Source |
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CRISPR system stocks for site-specific mutagenesis | The Bloomington Stock Center's CRISPR/Cas9 page. Includes various stocks for expression of both Cas9 and tracrRNA. | Bloomington Stock Center Indiana University Indiana, USA |
CRISPR Fly Design | Stocks available from the CRISPR Fly Design group at the MRC Laboratory of Molecular Biology. | MRC Laboratory of Molecular Biology Cambridge, UK |
CRISPR Vectors
Resource | Description | Author/Source |
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Addgene CRISPR/Cas9 Plasmids for use in insects | A list of available Cas9 & gRNA expression plasmids for use in insects, including Drosophila. | Addgene Cambridge, MA, USA |
CRISPR Fly Design Plasmids | Descriptions and information regarding the plasmids created by the CRISPR Fly Design group (also deposited at Addgene). | MRC Laboratory of Molecular Biology Cambridge, UK |
Additional Useful Links
Resource | Description | Author/Source |
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CRISPR Fly Design | An overview and resource collection for CRISPR gene editing in Drosophila. | MRC Laboratory of Molecular Biology Cambridge, UK |
flyCRISPR | An overview and resource collection for CRISPR gene editing in Drosophila. | O'Connor-Giles, Wildonger, and Harrison Labs University of Wisconsin-Madison WI, USA |
Insect Genomic Technologies Research Coordination Network | Discussion and dissemination of advanced technologies for genome modification of insects through symposia, technical workshops, and training fellowships. | Institute for Bioscience and Biotechnology Research University of Maryland Rockville, MD, USA |
FlyRNAi Blog, CRISPR tags | A research blog tracking functional genomics techniques in Drosophila, filtered for keyword: CRISPRs. | DRSC Harvard Medical School Boston, MA, USA |
DRSC Single Cell Cloning Protocol | Single-cell cloning method useful for Drosophila cell CRISPR gene editing | DRSC Harvard Medical School Boston, MA, USA |
Selected recent methods reviews
- Venken KJ, Sarrion-Perdigones A, Vandeventer PJ, Abel NS, Christiansen AE, Hoffman KL. Genome engineering: Drosophila melanogaster and beyond. Wiley Interdiscip Rev Dev Biol. 2015 Oct 8. doi: 10.1002/wdev.214. [Epub ahead of print] Review. PubMed PMID: 26447401.
- Beumer KJ, Carroll D. Targeted genome engineering techniques in Drosophila. Methods. 2014 Jun 15;68(1):29-37. doi: 10.1016/j.ymeth.2013.12.002. Epub 2014 Jan 8. Review. PubMed PMID: 24412316; PubMed Central PMCID: PMC4048800.
- Xu J, Ren X, Sun J, Wang X, Qiao HH, Xu BW, Liu LP, Ni JQ. A Toolkit of CRISPR-Based Genome Editing Systems in Drosophila. J Genet Genomics. 2015 Apr 20;42(4):141-9. doi: 10.1016/j.jgg.2015.02.007. Epub 2015 Mar 12. Review. PubMed PMID: 25953352.
Selected recent methods reports for tissue culture cells
- Bassett AR, Tibbit C, Ponting CP, Liu JL. Mutagenesis and homologous recombination in Drosophila cell lines using CRISPR/Cas9. Biol Open. 2014 Jan 15;3(1):42-9. doi: 10.1242/bio.20137120. PMID: 24326186.
- Böttcher R, Hollmann M, Merk K, Nitschko V, Obermaier C, Philippou-Massier J, Wieland I, Gaul U, Förstemann K. Efficient chromosomal gene modification with CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells. Nucleic Acids Res. 2014 Jun;42(11):e89. doi: 10.1093/nar/gku289. Epub 2014 Apr 19. PMID: 24748663
- Bassett AR, Kong L, Liu JL. A genome-wide CRISPR library for high-throughput genetic screening in Drosophila cells. J Genet Genomics. 2015 Jun 20;42(6):301-9. doi: 10.1016/j.jgg.2015.03.011. Epub 2015 Apr 18. PMID: 26165496
- Housden BE, Valvezan AJ, Kelley C, Sopko R, Hu Y, Roesel C, Lin S, Buckner M, Tao R, Yilmazel B, Mohr SE, Manning BD, Perrimon N. Identification of potential drug targets for tuberous sclerosis complex by synthetic screens combining CRISPR-based knockouts with RNAi. Sci Signal. 2015 Sep 8;8(393):rs9. doi: 10.1126/scisignal.aab3729. PMID: 26350902
Selected recent methods reports for in vivo
- Gratz SJ, Cummings AM, Nguyen JN, Hamm DC, Donohue LK, Harrison MM, Wildonger J, O'Connor-Giles KM. Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Genetics. 2013 Aug;194(4):1029-35. doi: 10.1534/genetics.113.152710. Epub 2013 May 24. PMID: 23709638
- Bassett AR, Tibbit C, Ponting CP, Liu JL. Highly efficient targeted mutagenesis of Drosophila with the CRISPR/Cas9 system. Cell Rep. 2013 Jul 11;4(1):220-8. doi: 10.1016/j.celrep.2013.06.020. Epub 2013 Jul 1. PMID: 23827738
- Yu Z, Ren M, Wang Z, Zhang B, Rong YS, Jiao R, Gao G. Highly efficient genome modifications mediated by CRISPR/Cas9 in Drosophila. Genetics. 2013 Sep;195(1):289-91. doi: 10.1534/genetics.113.153825. Epub 2013 Jul 5. PMID: 23833182
- Kondo S, Ueda R. Highly improved gene targeting by germline-specific Cas9 expression in Drosophila. Genetics. 2013 Nov;195(3):715-21. doi: 10.1534/genetics.113.156737. Epub 2013 Sep 3. PMID: 24002648
- Ren X, Sun J, Housden BE, Hu Y, Roesel C, Lin S, Liu LP, Yang Z, Mao D, Sun L, Wu Q, Ji JY, Xi J, Mohr SE, Xu J, Perrimon N, Ni JQ. Optimized gene editing technology for Drosophila melanogaster using germ line-specific Cas9. Proc Natl Acad Sci U S A. 2013 Nov 19;110(47):19012-7. doi: 10.1073/pnas.1318481110. Epub 2013 Nov 4. PMID: 24191015
- Port F, Muschalik N, Bullock SL. Systematic Evaluation of Drosophila CRISPR Tools Reveals Safe and Robust Alternatives to Autonomous Gene Drives in Basic Research. G3 (Bethesda). 2015 May 20;5(7):1493-502. doi: 10.1534/g3.115.019083. PMID: 25999583
- Ren X, Yang Z, Xu J, Sun J, Mao D, Hu Y, Yang SJ, Qiao HH, Wang X, Hu Q, Deng P, Liu LP, Ji JY, Li JB, Ni JQ. Enhanced specificity and efficiency of the CRISPR/Cas9 system with optimized sgRNA parameters in Drosophila. Cell Rep. 2014 Nov 6;9(3):1151-62. doi: 10.1016/j.celrep.2014.09.044. Epub 2014 Oct 23. PMID: 25437567
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RNAi
Introduction
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RNAi Design/Analysis Tools
Resource | Description | Author/Source |
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E-RNAi | E-RNAi is a tool for the design and evaluation of RNAi reagents. It can be used to design and evaluate long dsRNAs (including esiRNAs) as well as siRNAs. | Boutros lab, DKFZ, Heidelberg, Germany |
NEXT-RNAi | Software for the design and evaluation of genome-wide RNAi libraries which performs all steps from the prediction of specific and efficient RNAi target sites to the visualization of designed reagents in their genomic context. | Boutros lab, DFKZ, Heidelberg, Germany |
SnapDragon| | Cell-based RNAi reagent design | DRSC, Harvard Medical School, Boston, MA, USA |
RNAi Stocks for in vivo studies (fly stocks)
Resource | Description | Author/Source |
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UP-TORR, Updated Targets of RNAi Reagents | A lookup tool for searching BDSC, TRiP, VDRC and NIG_FLY in vivo RNAi collections. | DRSC, Harvard Medical School, Boston, MA, USA |
NIG_FLY | NIG-Fly in Mishima, Japan is part of a consortium with Kyoto-Fly and two other stock centers, and distributes RNAi flies on request. | NIG-FLY, Mishima, Japan |
BDSC RNAi Home Page | Links to lists and sublists of RNAi and miRNA sponge insertions at Bloomington. Most, but not all, of the insertions are under the control of UAS and therefore require GAL4 for expression. | BDSC, Bloomington Drosophila Stock Center, Bloomington, IN, USA |
TRiP Transgenic RNAi Project | The TRiP Stock Collection contains over 9,000 lines, 1,575 of which are for orthologs of human-disease-associated genes. The stocks can be obtained from the BDSC. | DRSC, Harvard Medical School, Boston, MA, USA |
THFC, Tsinghua Fly Center | THFC RNAi Stock Collection for triggering RNAi in soma and germline | Tsinghua Fly Center, Beijing, China |
VDRC | The VDRC at IMP/IMBA in Vienna provides two genome-wide transgenic Drosophila RNAi libraries | VDRC, Vienna, Austria |
Cell-based RNAi Reagents
Resource | Description | Author/Source |
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DRSC Libraries | Genome-Wide Libraries for cell-based RNAi Screening and Validation (on-site or shipped) | DRSC, Harvard Medical School, Boston, MA, USA |
Division of Signaling and Functional Genomics | Genome-Wide Libraries for cell-based RNAi Screening and Validation | Boutros lab, DKFZ, Heidelberg, Germany |
RNAi Screening Centers
Resource | Description | Author/Source |
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DRSC, Drosophila RNAi Screening Center | The Drosophila RNAi Screening Center at Harvard Medical School provides high throughput RNAi screens | DRSC, Harvard Medical School, Boston, MA, USA |
NYU RNAi Screening and Automation Core | The NYU RNAi Core Facility at the Langone Medical Center in New York provides high-throughput RNAi screens. Drosophila screens are offered in collaboration with the DRSC | NYU RNAi Core, New York, NY, USA |
Sheffield RNAi Screening Facility| | The SRSF provides a service for whole-genome RNAi screens in Drosophila cells. | SRSF, University of Sheffield, Boston, MA, USA |
RNAi Results, Validation and Phenotypes
Resource | Description | Author/Source |
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GenomeRNAi | GenomeRNAi is a database containing phenotypes from cell-based and in vivo RNA interference (RNAi) screens in Drosophila | GenomeRNAi, DKFZ, Heidelberg, Germany |
RNAi Stock Validation and Phenotypes for in vivo RNAi studies | DRSC, Harvard Medical School, Boston, MA, USA | |
Cell-Based Screen Summary | A summary of public cell-based DRSC screens | DRSC, Harvard Medical School, Boston, MA, USA |
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Stock Centers
List of Drosophila Stock Centers
Resource | Author/Source | Additional Information |
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CRISPR system stocks for site-specific mutagenesis | Bloomington Stock Center | Lorem ipsum dolor sit amet, consectetur adipiscing elit. Nullam et tincidunt ante. Donec tincidunt gravida velit at iaculis. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Aenean rutrum quam eget luctus condimentum. Cras in venenatis lectus, at laoreet augue. |
CRISPR Fly Design | MRC - Laboratory of Molecular Biology | Lorem ipsum dolor sit amet, consectetur adipiscing elit. Integer id purus metus. Cras et lacus vel diam posuere facilisis. Vestibulum lorem metus, dapibus nec porta non, fringilla sit amet eros. |
Example | Example | Example |
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Antibodies
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Additional Resources
For additional information, please see our extensive lists: