CRISPR/Cas Tools

This page describes a list of software platforms and bioinformatics tools built to facilitate the design of guide RNAs (gRNAs) for use with the CRISPR/Cas system.

The CRISPR/Cas (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated nucleases) system was originally discovered to be an acquired immune response mechanism used by archaea and bacteria. It has since been adopted for use as a tool in the genetic engineering of higher organisms.

Designing an appropriate gRNA is an important element of genome editing with the CRISPR/Cas system. A gRNA can and at times does have unintended interactions ("off-targets") with other locations of the genome of interest. For a given candidate gRNA, these tools report its list of potential off-targets in the genome thereby allowing the designer to evaluate its suitability prior to embarking on any experiments.

Scientists have also begun exploring the mechanics of the CRISPR/Cas system and what governs how good, or active, a gRNA is at directing the Cas nuclease to a specific location of the genome of interest.[1][2] As a result of this work, new methods of assessing a gRNA for its 'activity' have been published,[1][2] and it is now best practice to consider both the unintended interactions of a gRNA as well as the predicted activity of a gRNA at the design stage.

The below table lists available tools and their attributes, and includes links to the corresponding websites.

List of CRISPR/Cas tools
Tool Name Provider Searches whole genome for targets Returns all targets of genome Seed span and location can be defined Maximum number of mismatches supported Predicts gRNA activity Available Protospacer adjacent motif (PAM) sequences Annotation is reported gRNA suggestion or scoring External Link References
Benchling CRISPR gRNA Design Benchling Yes Yes Yes 4 Yes User customizable Yes Yes Webserver -
Breaking-Cas Spanish National Center for Biotechnology Yes Yes Yes (by weights) 4 No User customizable Yes Yes Webserver [3]
Cas-OFFinder Seoul National University Yes Yes No 0-10 No NGG, NRG, NNAGAAW, NNNNGMTT No Yes Webserver Source code [4]
CCTop University of Heidelberg Yes Yes Partial 5 (0-5) No NGG, NRG, NNGRRT, NNNNGATT, NNAGAAW, NAAAAC Yes Yes Webserver [5]
CHOPCHOP Harvard University Yes Yes Partial 0, 2 No NGG, NNAGAA, NNNNGANN No Yes Webserver [6]
CHOPCHOP v2 University of Bergen Yes Yes Yes 3 (0-3) Yes User customizable Yes Yes Webserver [7]
COD Dayong Guo No No No 0, 3, 5, 8 No NGG and NAG No Yes Webserver -
CRISPR Configurator & Specificity Tool Dharmacon, Inc. Yes (over 30 species) Yes Yes 8 (gaps or mismatches) Internally NGG and NAG mRNA exons, Links to UCSC genome browser annotations No Webserver Specificity Tool -
CRISPR Design Zhang Lab, MIT Yes No No 4 No NGG and NAG mRNA exons Yes Webserver [8]
CRISPRdirect Database Center for Life Science (DBCLS) Yes (over 200 species) Yes No Any number No NNN Yes Yes Webserver [9]
CRISPR gRNA Design Tool DNA2.0 Yes Yes No 0-10 No NGG, NAG Genbank annotations: Gene, misc_RNA, ncRNA, CDS, exon Yes Webserver -
CRISPRseek Bioconductor Yes Yes No Any number No User customizable mRNA exons Yes Source code [10]
DESKGEN Desktop Genetics Yes Yes Yes Any number Yes Fully user customizable Yes Yes Webserver [11]
Off-Spotter Thomas Jefferson University Yes Yes Yes 0-5 NGG, NAG, NNNNACA, NNGRRT (R is A or G) mRNA exons, unspliced mRNA, mRNA, 5'UTR, CDS, 3'UTR, unspliced lincRNA, lincRNA User customizable Webserver Source code Remote batch submission[12]
sgRNA Designer Broad Institute No No No 0 Yes NGG CDS (if searching by transcript ID) Yes Webserver Source code [1]

References

  1. 1 2 3 Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I, Sullender M, Ebert BL, Xavier RJ, Root DE (2014). "Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation". Nat. Biotechnol. 32 (12): 1262–7. doi:10.1038/nbt.3026. PMID 25184501.
  2. 1 2 Chari, Raj; Mali, Prashant; Moosburner, Mark; Church, George M (2015-01-01). "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach". Nature Methods. 12: 823–826. doi:10.1038/nmeth.3473.
  3. ↑ Oliveros, Juan C.; Franch, Mònica; Tabas-Madrid, Daniel; San-León, David; Montoliu, Lluis; Cubas, Pilar; Pazos, Florencio (2016-07-08). "Breaking-Cas-interactive design of guide RNAs for CRISPR-Cas experiments for ENSEMBL genomes". Nucleic Acids Research. 44 (W1): W267–271. doi:10.1093/nar/gkw407. ISSN 1362-4962. PMC 4987939Freely accessible. PMID 27166368.
  4. ↑ Bae S, Park J, Kim JS (2014). "Cas-OFFinder: a fast and versatile algorithm that searches for potential off-target sites of Cas9 RNA-guided endonucleases". Bioinformatics. 30 (10): 1473–5. doi:10.1093/bioinformatics/btu048. PMID 24463181.
  5. ↑ Stemmer M, Thumberger T, Del Sol Keyer M, Wittbrodt J, Mateo JL (2015). "CCTop: An Intuitive, Flexible and Reliable CRISPR/Cas9 Target Prediction Tool". PLOS ONE. 10: e0124633. doi:10.1371/journal.pone.0124633. PMC 4409221Freely accessible. PMID 25909470.
  6. ↑ Montague TG, Cruz JM, Gagnon JA, Church GM, Valen E (2014). "CHOPCHOP: a CRISPR/Cas9 and TALEN web tool for genome editing". Nucleic Acids Res. 42 (W1): W401–7. doi:10.1093/nar/gku410. PMID 24861617.
  7. ↑ Labun K, Montague TG, Gagnon JA, Thyme SB, Valen E (2016). "CHOPCHOP v2: a web tool for the next generation of CRISPR genome engineering.". Nucleic Acids Res. 44 (W1): W272–6. doi:10.1093/nar/gkw398. PMC 4987937Freely accessible. PMID 27185894.
  8. ↑ Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O, Cradick TJ, Marraffini LA, Bao G, Zhang F (2013). "DNA targeting specificity of RNA-guided Cas9 nucleases". Nat. Biotechnol. 31 (9): 827–32. doi:10.1038/nbt.2647. PMID 23873081.
  9. ↑ Naito Y, Hino K, Bono H, Ui-Tei K (2015). "CRISPRdirect: software for designing CRISPR/Cas guide RNA with reduced off-target sites". Bioinformatics. 31 (7): 1120–3. doi:10.1093/bioinformatics/btu743. PMC 4382898Freely accessible. PMID 25414360.
  10. ↑ Zhu LJ, Holmes BR, Aronin N, Brodsky MH (2014). "CRISPRseek: A Bioconductor Package to Identify Target-Specific Guide RNAs for CRISPR-Cas9 Genome-Editing Systems". PLOS ONE. 9 (9): e108424. doi:10.1371/journal.pone.0108424. PMC 4172692Freely accessible. PMID 25247697.
  11. ↑ "Desktop Genetics Announces the Launch of DeskGen Gene Editing Platform". American Laboratory. 2015.
  12. ↑ Pliatsika V, Rigoutsos I (2015). ""Off-Spotter": very fast and exhaustive enumeration of genomic lookalikes for designing CRISPR/ Cas guide RNAs". Biol. Direct. 10 (1): 4. doi:10.1186/s13062-015-0035-z. PMID 25630343.

External links

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