CRISPR/Cas Tools

CRISPR/Cas Tools

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated (Cas) proteins perform adaptive immune functions in prokaryotic organisms defending against foreign nucleic acids such as viruses. CRISPR/Cas tools have been adapted for use in genome editing and other DNA and RNA targeting applications.

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Edits on 27 Feb, 2020
Meredith Hanel
Meredith Hanel edited on 27 Feb, 2020
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  • Cas9 and Cas12 based systems by Caspr Biotech to detect viruses, bacteria and genetic mutations
Edits on 17 Sep, 2019
Meredith Hanel
Meredith Hanel edited on 17 Sep, 2019
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  • Programmed chromosome fissionProgrammed chromosome fission and fusionfusion in E. coli with Cas9
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RecombineeringRecombineering is a method of genetic engineering in bacterial genomes that involves homologous recombination. In contrast to CRISPR used for gene editing in eukaryotic cells, CRISPR assists the selection of successful recombineering events which change the target sequence and negatively selects cells in which the desired homologous recombination has not occurred. Negative selection occcurs by CRISPR-Cas9 induced double stranded breaks which are lethal in bacteria where non homologous end joining (NHEJ) is not very effective.

Edits on 16 Sep, 2019
Meredith Hanel
Meredith Hanel edited on 16 Sep, 2019
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  • Programmed chromosome fission and fusion in E. coli with Cas9
Edits on 20 Jun, 2019
Meredith Hanel
Meredith Hanel edited on 20 Jun, 2019
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Edits on 27 Mar, 2019
Meredith Hanel
Meredith Hanel edited on 27 Mar, 2019
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  • CRISPR-Chip
Edits on 22 Feb, 2019
Meredith Hanel
Meredith Hanel edited on 22 Feb, 2019
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  • CRISPR/Cas9 insertion of MS2 casette for RNA tracking. MS2 labeling of RNA is a method that tags an RNA of interest with a stem-loop RNA sequence derived from the bacteriophage MS2 genome which binds to bacteriophage coat protein. The coat protein is fused to a fluorescent protein so that it can be visualized and it binds to the stem-loop tagged RNA.
Edits on 6 Feb, 2019
Meredith Hanel
Meredith Hanel edited on 6 Feb, 2019
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Gene/genome editing therapeutics companies

Edits on 21 Jan, 2019
Meredith Hanel
Meredith Hanel edited on 21 Jan, 2019
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CRISPR-Cas systems are being developedcombined with phage therapy to target and degrade DNA of pathogenic bacteria as a selective antimicrobial treatment

Edits on 19 Jan, 2019
Meredith Hanel
Meredith Hanel edited on 19 Jan, 2019
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  • Nextbiotics
Edits on 14 Jan, 2019
Meredith Hanel
Meredith Hanel edited on 14 Jan, 2019
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CRISPR-Cas3CRISPR-Cas issystems are being developed by Locus Biosciences to target and degrade the DNA of pathogenic bacteria in orderas toa killselective it.antimicrobial treatment

  • Locus Biosciences (CRISPR-Cas3)
  • Eligo Bioscience (CRISPR-Cas9)
Edits on 13 Dec, 2018
Meredith Hanel
Meredith Hanel edited on 13 Dec, 2018
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  • Cas13a based SHERLOCKSHERLOCK to detect pathogenic viruses and bacteria
Edits on 5 Nov, 2018
Meredith Hanel
Meredith Hanel edited on 5 Nov, 2018
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  • MAGESTIC (Multiplexed Accurate Genome Editing with Short, Trackable, Integrated Cellular barcodes) which can be used to study how sequence variants impact cell phenotypes on a large scale
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Genome editing with CRISPR-Cas9 has been demonstrated in model species such as Arabidopsis, rice and tobacco as well as a few crop species. CRISPR-Cas12a genome editing has also been demonstrated to work in plants. The United States Department of Agriculture (USDA) stated that CRISPR-Cas9 edited crops will not be regulated as GMO.

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  • Waxy corn enriched with amylopectin due to inactivation of endogenous waxy gene
Edits on 5 Nov, 2018
Meredith Hanel
Meredith Hanel edited on 5 Nov, 2018
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Related Topics (+1 topics)
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  • Cas12a
  • Cas12a based DETECTR system to detect viruses
  • Cas12a based HOMES system to detect viruses and small nucleotide polymorphism sites in human DNA related to health and personal characteristics
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Based on the ability of CRISPR systems to sequentially acquire DNA sequences from viral infections, adding them to spacer sequences in a growing array in the CRISPR locus, molecular recorder tools are being developed which can record information provided by researchers or from the cell interactions with the environment.

  • DNA based storage. Cas1 and Cas2 have been engineered by researchers at the Wyss Institute and Harvard Medical School to record pixel information in DNA so that the information could be retrieved to reconstruct frames in a galloping horse movie.
  • CRISPR-Cas RNA recording system. A tool to record transient events where genes are active and transcribing RNA and record them in DNA. In order to accomplish this researchers at ETH Zurich, Switzerland fused a Cas gene to reverse transcriptase, which copies RNA into DNA.

Related Topics
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
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  • Cas9 nickase, where Cas9 is modified to produce a single strand break and CRISPR-Cas9 nickases are delivered as pairs for more target specificity
  • photoactivated Cas9 (paCas9) for optogenetic genome editing
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CRISPR knock outlibrary screening

Genetic screening of mutant libraries is a way to search for genes involved in a desired pathway where mutant genes produce a certain phenotype. CRISPR libraries contain thousands of plasmids with multiple guide RNAs (gRNAs) for each target gene. Cells are treated with the library gRNA and Cas9 creating a population of mutant cells which can be screened for a phenotype of interest. CRISPR libraries can knockout, activate or repress target genes. The following companies and non-profits sell or distribute CRISPR libraries.

  • Twist Bioscience
  • ThermoFisher Scientific
  • Addgene (non-profit plasmid repository)
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  • anti-CRISPR protein AcrIIA4 and LOV photosensor for optogenetic control of CRISPR-Cas9
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
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  • Cas9 nickase
  • Cas9 nickase, where Cas9 is modified to produce a single strand break and CRISPR-Cas9 nickases are delivered as pairs for more target specificity
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
Edits made to:
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Genome editing with CRISPR-Cas9 has been demonstrated in model species such as Arabidopsis, rice and tobacco as well as a few crop species. CRISPR-Cas12a genome editing has also been demonstrated to work in plants. The United States Department of Agriculture (USDA) stated that CRISPR-Cas9 edited crops will not be regulated as GMO.

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The following crops have been improved using CRISPR-Cas9 technology:

  • Non-browning white button mushroom where polyphenol oxidase is knocked out
  • Waxy corn enriched with amylopectin due to inactivation of endogenous waxy gene
  • Green bristlegrass with delayed flowering due to inactivation of the ID1 gene
  • Camelina with increased oil content by Yield10 Bioscience
  • Drought tolerant soybean with edited Drb2a and Drb2b genes
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
Edits made to:
Article (+754 characters)
Article

Genome editing with CRISPR-Cas9 has been demonstrated in model species such as Arabidopsis, rice and tobacco as well as a few crop species. CRISPR-Cas12a genome editing has also been demonstrated to work in plants. The United States Department of Agriculture (USDA) stated that CRISPR-Cas9 edited crops will not be regulated as GMO.

The following crops have been improved using CRISPR-Cas9 technology:

  • Non-browning white button mushroom where polyphenol oxidase is knocked out
  • Waxy corn enriched with amylopectin due to inactivation of endogenous waxy gene
  • Green bristlegrass with delayed flowering due to inactivation of the ID1 gene
  • Camelina with increased oil content by Yield10 Bioscience
  • Drought tolerant soybean with edited Drb2a and Drb2b genes

Edits on 4 Nov, 2018
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
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Agriculture
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
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CRISPR-Cas molecular recording
Meredith Hanel
Meredith Hanel edited on 4 Nov, 2018
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  • dCas9-DNMT3A
  • dCas9-Ezh2
  • dCas9-DNMT3B
  • dCas9-TET1/2
  • dCas9-KRAB
  • dCas9-MQ1 (prokaryotic DNA methyltransferase)
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The organization of genes within the genome and their location within chromatin architecture such as chromatin loop structures is important for gene regulation programs in development and disease.

  • CLOuD9 is a dCas9 based system which selectively and reversibly establishes chromatin loops
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Anti-CRISPR proteins

Anti-CRISPR proteins are a defense mechanism which evolved in phages to escape destruction from CRISPR in their host prokaryotes. Anti-CRISPR proteins have applications as an “off switch” for CRISPR based genome editing or for reducing off-target cutting by CRISPR systems.

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