Genome Editing CRISPR Cas9

Genome Editing

Genome editing is enabled by the development of tools to make precise, targeted changes to the genome of living cells. Recently a new tool based on a bacterial CRISPR-associated protein-9 nuclease (Cas9) from Streptococcus pyogenes has generated considerable excitement. This follows several attempts over the years to manipulate gene function, including homologous recombination and RNA interference. RNAi, in particular, became a laboratory staple enabling inexpensive and high-throughput interrogation of gene function, but is hampered by providing only temporary inhibition of gene function and unpredictable off-target effects. Other recent approaches to targeted genome modification — zinc-finger nucleases [ZFNs] and transcription-activator like effector nucleases [TALENs]– enable researchers to generate mutations by introducing double-stranded breaks to activate repair pathways. These approaches are costly and time consuming to engineer, limiting their widespread use, particularly for large scale, high-throughput studies.

CRISPR (Clustered Regulary Interspaced Short Palindromic Repeats) and CRISPR-associated (Cas) genes are essential in adaptive immunity in select bacteria and archaea, enabling the organisms to respond to and eliminate invading genetic material.

The simplicity of the CRISPR nuclease system, with only three components (Cas9 , crRNA and trRNA) makes this system amenable to adaptation for genome editing. By combining the crRNA and trRNA into a single synthetic guide RNA (sgRNA), a further simplified two component system can be used to introduce a targeted double stranded break. This break activates repair through error prone non-homologous end joining (NHEJ) or Homology directed Repair (HDR). In the presence of a donor template with homology to the targeted locus, the HDR pathway operates allowing for precise mutations to be made. In the absence of a template, NHEJ is activated resulting in insertions and/or deletions (indels) which disrupt the target locus.

Genome engineering with Cas9 Nuclease

NEB TV – Episode 7

Learn more about CRISPR/Cas9 and synthetic biology, where these fields are headed, and how they are being applied in the classroom in the latest episode of NEB TV.

Cas9 in vivo: Bacterial Adaptive Immunity

CRISPR systems evolved as a means of bacterial adaptive immunity. Understanding their behavior in vivo is essential to harnessing their power in vitro.

Selected products for CRISPR/Cas9 Workflow

CRISPR/Cas9 Application PRODUCT Prod.Nr.: SIZE PRICE*
Central Component of the CRISPR/Cas9-Workflows EnGen® Spy Cas9 NLS M0646T 400 pmol 99 €* In den Shop wechseln Produktinformationen
M0646M 2000 pmol 320 €*
Central Component of the CRISPR/Cas9-Workflows, without NLS Cas9 Nuclease,
S. pyogenes
M0386S 70 pmol 41 €* In den Shop wechseln Produktinformationen
M0386T 400 pmol 99 €*
M0386M 2000 pmol 320 €*
  • Editing in AT rich regions
  • Maintains activity at lower temperatures than the Acidaminococcus orthologs, permitting editing in ectothermic organisms such as zebra fish and xenopus
EnGen® Lba Cas12a (Cpf1) M0653S 70 pmol 74 € In den Shop wechseln Produktinformationen
M0653T 2000 pmol 264 €
Paired, offset cleavage of genomic loci. Target-specific DNA nicking EnGen® Spy Cas9 Nickase M0650S 70 pmol 45 €* In den Shop wechseln Produktinformationen
M0650T 400 pmol 99 €*
  • Localization of DNA targets in vivo
  • Target enrichment
EnGen®Spy dCas9 (SNAP-tag®) M0652S 70 pmol 45 €* In den Shop wechseln Produktinformationen
M0652T 400 pmol 99 €*
Synthesis of sgRNA EnGen® sgRNA Synthesis Kit,S. pyogenes E3322S 20 rxns 299 €* In den Shop wechseln Produktinformationen
Detection of on-target genome editing events EnGen® Mutation Detection Kit E3321S 25 rxns 210 € In den Shop wechseln Produktinformationen
Additionally available NEB products for the use in CRISPR Workflows:
Generation of sgRNA HiScribe T7 High Yield
RNA Synthesis Kit
E2040S 50 rxns 223 € In den Shop wechseln Produktinformationen
HiScribe T7 Quick High Yield RNA Synthesis Kit E2050S 50 rxns 273 € In den Shop wechseln Produktinformationen
Generation of capped Cas9 mRNA HiScribe T7 ARCA mRNA Kit (with Tailing) E2060S 20 rxns 376 € In den Shop wechseln Produktinformationen
HiScribe T7 ARCA mRNA Kit E2065S 20 rxns 322 € In den Shop wechseln Produktinformationen
Insertion of target sequence into the Cas9-sgRNA construct Q5 Site-directed
Mutagenesis Kit (with or without comp. E. coli)
E0554S 10 rxns
(+ komp. E.coli)
192 € In den Shop wechseln Produktinformationen
E0552S 10 rxns 132 € In den Shop wechseln Produktinformationen
High-fidelity construct generation for use with CRISPR workflows Q5 High-Fidelity
DNA Polymerase
M0491S 100 u 96 € In den Shop wechseln Produktinformationen
M0491L 500 u 424 €
Q5 Hot Start High-Fidelity DNA Polymerase M0493S 100 u 112 € In den Shop wechseln Produktinformationen
M0493L 500 u 508 €
Single-tube, isothermal generation of the Cas9-sgRNA construct NEBuilder HiFi DNA Assembly Master Mix E2621S 10 rxns 162 € In den Shop wechseln Produktinformationen
E2621L 50 rxns 648 €
E2621X 250 rxns 2.589 €
NEBuilder HiFi DNA Assembly Cloning Kit
(incl. comp. E. coli)
E5520S 10 rxns 190 € In den Shop wechseln Produktinformationen

As of: 03.01.2019

*Permanent low prices – no further discounts apply. Offer closes 31.12.2019.

Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.