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CRISPR header icon

Multiplex CRISPR dCas9/FokI-dCas9 Accessory Pack
(Kit # 1000000062 )

Depositing Lab: Takashi Yamamoto

This supplemental package contains plasmids for dCas9 and FokI-dCas9 expression used to construct all-in-one CRISPR/Cas9 vectors expressing multiple gRNAs.

This kit consists of 12 individually packaged plasmids, which are shipped as bacterial stabs at room temperature. Please note that the Multiplex CRISPR/Cas9 Assembly System Kit is required to use the plasmids in this accessory pack for multiplex gRNA assembly.

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$375 USD + shipping

Available to academics and nonprofits only.

Original Publication

Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T. BMC Biotechnol. 2015 May 22;15(1):33. doi:10.1186/s12896-015-0144-x. PubMed .

Description

The Multiplex CRISPR dCas9/FokI-dCas9 Accessory Pack enables construction of all-in-one CRISPR/Cas9 vectors expressing multiple gRNAs (2-7) and dCas9 or dCas9-FokI fusion . Multiple gRNA cassettes are assembled using BsaI-mediated Golden Gate cloning method. The kit contains 12 plasmids: pX330A_dCas9-1x[2-7] and pX330A_FokI-1x[2-7].

Overview of Multiplex CRISPR dCas9/FokI-dCas9 plasmids

Figure illustrating assembly process using dCas9 or FokI/dCas9 accessory plasmids

Figure adapted from Nakagawa et al., BMC Biotechnol. 2015 May 22;15(1):33.

Protocol

Download a PDF Protocol for Multiplex CRISPR construction as described in the above article:

Icon Multiplex CRISPR Kit assembly protocol (365.9 KB)

Resources

Download a ZIP file containing all Multiplex CRISPR dCas9/FokI-dCas9 Accessory Pack plasmid sequences in GenBank format:

Icon Annotated GenBank files (88.9 KB)

How to Cite this Kit

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods of your future publications.

For your Materials and Methods section:
"The CRISPR kit used for constructing multiplex CRISPR/Cas9 vectors was a gift from Takashi Yamamoto (Addgene kit # 1000000062)"

For your Reference section:
Production of knockout mice by DNA microinjection of various CRISPR/Cas9 vectors into freeze-thawed fertilized oocytes. Nakagawa Y, Sakuma T, Sakamoto T, Ohmuraya M, Nakagata N, Yamamoto T. BMC Biotechnol. 2015 May 22;15(1):33. doi:10.1186/s12896-015-0144-x. PubMed .

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