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CRISPR/Cas9 Genome Editing: pDNA + gRNA Transfection

TransIT-X2® Dynamic Delivery System for Plasmid DNA and Guide RNA Oligonucleotide Delivery

Genome Editing

  • Cas9 protein and guide RNA can both be encoded as plasmid DNA for transfection. Alternatively, Cas9 can be delivered as plasmid DNA, and guide RNA can be supplied as an RNA oligonucleotide.

Benefits to these approaches include:

  • Low Cost – Plasmid DNA is a renewable, cost-effective format
  • Flexibility – Cas9 and guide RNA plasmids are suitable for stable or transient transfection
  • Ease of use – Guide RNA oligonucleotide format enables simple retargeting of Cas9 to different loci

CRISPR/Cas9 Delivery Methods - Cas9 and Guide RNA Plasmids

CRISPR/Cas9 Plasmid Delivery Approaches

CRISPR/Cas9 Delivery Methods – Cas9 and Guide RNA Plasmids.
(A) Cas9 and guide RNA are encoded on the same plasmid. (B,C) Cas9 and guide RNA(s) are encoded on separate plasmids. (A,B) The wild-type Cas9 enzyme contains two endonuclease domains which cleave the target DNA on both strands when programmed with a guide RNA. (C) The D10A mutation converts Cas9 into a nickase that generates single-stranded breaks in the target DNA. For improved target specificity, Cas9 D10A can be used with paired guide RNAs targeting opposite strands to create staggered double-stranded breaks.

CRISPR/Cas9 Delivery Methods - Cas9 Plasmid + Guide RNA Oligonucleotides

CRISPR/Cas9 Plasmid and Guide RNA Delivery Approaches

CRISPR/Cas9 Delivery Methods – Cas9 Plasmid + Guide RNA Oligonucleotides.
Cas9 is supplied as plasmid DNA, and guide RNA(s) are supplied as either synthetic or in vitro transcribed RNA oligonucleotides. (A) The wild-type Cas9 enzyme contains two endonuclease domains which cleave the target DNA on both strands when programmed with a guide RNA. (B) The D10A mutation converts Cas9 into a nickase that generates single-stranded breaks in the target DNA. For improved target specificity, Cas9 D10A can be used with paired guide RNAs targeting opposite strands to create staggered double-stranded breaks.

Efficient Genome Editing with Cas9 Plasmid DNA and Guide RNA Oligonucleotides

CRISPR Plasmid and gRNA Delivery Data

Efficient Genome Editing with Cas9 Plasmid DNA + Guide RNA Oligonucleotides.
HEK293T/17, U2OS, and NHDF cells were co-transfected with 0.5 µg of Cas9-encoding pDNA (MilliporeSigma) and 50 nM PPIB targeting 2-part gRNA (Dharmacon) using TransIT-X2 Dynamic Delivery System (2 µl/well of a 24-well plate, Mirus Bio). A T7E1 mismatch detection assay was used to measure cleavage efficiency at 48 hours post-transfection.

ORDER INFORMATION

  • For shipping and storage information please click on Order#.
Order# Description Amount Price Data Sheet
MIR6000 TransIT-X2® Dynamic Delivery System 1.5 ml 547,00 PDF
MIR6003 TransIT-X2® Dynamic Delivery System 0.3 ml 132,00 PDF
MIR6004 TransIT-X2® Dynamic Delivery System 0.75 ml 329,00 PDF
MIR6005 TransIT-X2® Dynamic Delivery System 5 x 1.5 ml 2374,00 PDF
MIR6006 TransIT-X2® Dynamic Delivery System 10 x 1.5 ml 4365,00 PDF
  • All prices are in EURO excl. VAT and shipping. Only available in selected countries.

  • * Sample Policy:
    MoBiTec together with Mirus considers sample requests for up to three different transfection products and one electroporation product per laboratory. Please include all samples to be tested in a single request.

Download

  • CRISPR-Cas9-Poster
  • Mirus CRISPR/Cas9 Transfection Poster (PDF):
    Optimization of DNA, RNA and RNP delivery methods for efficient CRISPR/Cas9 Mediated Mammalian Cell Engineering