Custom CRISPR Knock-in Repair Templates

From a single-nucleotide substitution to a fluorescent reporter tag - a CRISPR knock-in creates precise desired mutations in the genomic DNA near the site where sequence-specific hybridization of an sgRNA recruits the Cas9 nuclease. The desired sequence is delivered into the cell as an HDR template (either an ssDNA oligonucleotide or a donor plasmid), which allows for homology-directed repair (HDR) to occur after Cas9 has made a double-stranded break in the genomic DNA. Every CRISPR HDR template consists of the desired mutation or insert sequence flanked by left and right homology arms that correspond to the sequence on either side of the CRISPR-induced double stranded break. For small mutations of <50 bp, we suggest using ssDNA oligonucleotide templates, while larger insertions should be handled with one of our dsDNA donor plasmids. Choose to use our Custom CRISPR HDR templates to achieve precise genomic knock-in using the homology-directed repair pathway.



Applications
  1. In-frame fusion tag
    • →Tag your gene of interest with a short peptide sequence
    • →For this application, an ssDNA oligonucleotide repair template is recommended
  2. Point mutation
    • →Mutate short sequences or residues within your gene of interest
    • →For this application, an ssDNA oligonucleotide repair template is recommended
  3. Transgene insertion
    • →Insert a larger transgene or reporter into a locus of your choice
    • →For this application, a dsDNA vector repair template is recommended
Testimonial:

"Construct works beautifully, definitely will get more if needed."

Emily Chen, UCLA, CRISPR Constructs

Additional Resources:

Service Details

ssDNA oligonucleotide service
Service NameHomology Arms + Repair TemplateQuantityCat. No.Price
Custom CRISPR Repair Template (<200nt ssDNA oligonucleotide) Homology arms + insert must be less than 200nt total. Suggested homology arm size is 50nt-90nt per arm 4 nmole C427 $275.00
  • Lyophilized oligonucleotide containing your insert with flanking homology arms
dsDNA Vectors with Repair Template
Service NameRepair InsertQuantityCat. No.Price
Custom CRISPR Repair Template (< 1.2 kbp arms, < 1 kbp insert) Custom insert < 1 kbp 1.0 ug C428 $615.00
Custom CRISPR Repair Template (< 1.2 kbp arms, 1-2 kbp insert) Custom insert 1-2 kbp 1.0 ug C429 $795.00
Custom CRISPR Repair Template (< 1.2 kbp arms, SV40-GFP-2A-Puro insert) Predesigned insert with GFP and Puromycin resistance 1.0 ug C430 $875.00
Custom CRISPR Repair Template (< 1.2 kbp arms, SV40-RFP-2A-Puro insert) Predesigned insert with GFP and Puromycin resistance 1.0 ug C431 $875.00
  • Plasmid with Homology Arms up to <1.2kb (both left and right arms)
  • dsDNA vector containing an insert with flanking homology arms (total homology arms sequence is < 1.2kb in length)
Plasmid with Homology Arms 1.2-2 kbp (total of both arms)
Service NameRepair InsertQuantityCat. No.Price
Custom CRISPR Repair Template (1.2-2 kbp arms, < 1 kbp insert) Custom insert < 1 kbp 1.0 ug C432 $855.00
Custom CRISPR Repair Template (1.2-2 kbp arms, 1-2 kbp insert) Custom insert 1-2 kbp 1.0 ug C433 $1025.00
Custom CRISPR Repair Template (1.2-2 kbp arms, SV40-GFP-2A-Puro insert) Predesigned with GFP and Puromycin resistance 1.0 ug C434 $1095.00
Custom CRISPR Repair Template (1.2-2 kbp arms, SV40-RFP-2A-Puro insert) Predesigned with GFP and Puromycin resistance 1.0 ug C435 $1095.00
  • dsDNA vector containing an insert with flanking homology arms (total homology arms sequence is 1.2-2kb in length)
Plasmid with Homology Arms >2 kbp (total of both arms)
Service NameRepair InsertQuantityCat. No.Price
Custom CRISPR Repair Template (>2 kbp arms, any insert size) Custom or predesigned insert 1.0 ug C436 Inquire
  • dsDNA vector containing an insert with flanking homology arms (total homology arms sequence is >2kb in length)

Additional Info

Workflow

Available custom CRISPR Vectors and Viruses:
AAV Vector/VirusLentivector/virusNon-Viral VectorAdenovirus
sgRNA for spCas9 Available Available Available Available
sgRNA for saCas9 Available - - -
All-In-One with spCas9 - Available Available -
All-In-One with saCas9 Available - - -

FAQs

Coming Soon

Citations

01 Kang, Y. J. et al. "Regulation of NKT cell-mediated immune responses to tumours and liver inflammation by mitochondrial PGAM5-Drp1 signalling." Nat. Commun. (2015) 6:8371 doi: 10.1038/ncomms9371
02 Jiang, G. et al. "Isorhapontigenin (ISO) inhibits invasive bladder cancer (BC) formation in vivo and human BC invasion in vitro by targeting STAT1/FOXO1 Axis." Cancer Prev Res. Published Online First April 14, 2016.doi: 10.1158/1940-6207.CAPR-15-0338
03 Okugawa, Y. et al. "Clinical significance of SNORA42 as an oncogene and a prognostic biomarker in colorectal cancer." Gut (2015)doi:10.1136/gutjnl-2015-309359