CUSTOM SERVICES

 

Custom shRNA Cloning

If you wish to use selection other than puromycin resistance, we can clone shRNA into one of the following three plasmids and provide sequence-verified plasmid constructs:

pLKO.1-puro-GFP

pLKO.1-puro-RFP

pLKO.1-neo-GFP

 

Turnaround time: 3-4 weeks

Cost: See Product prices (Contact us for large orders)

 

Custom shRNA Panel

You provide a list of genes and the number of shRNA per gene you wish to include in the custom panel. We generate a shRNA list and pool them from the TRC library. Individual bacterial stocks are cultured overnight in 96-well deep well plates, pooled and plasmid isolated using Qiagen Maxi Plus kits. Backup glycerol plates are also generated for every custom 96-well plate. Finally, all plasmid preps are pooled in equimolar ratio. This pooled plasmid prep is provided provided to you as plasmid DNA or lentiviral suspension.

 

Turn around time: Variable

Cost: See Product prices (Contact us for large panels)

 

Custom gRNA Cloning

If you wish to use an all-in-one system to simultaneously deliver gRNA and Cas9, we can clone gRNA into the following vectors:

PX458

PX459

PX459v2

PX461

PX462

lentiCRISPRv2-puro

lentiCRISPRv2-blast

lentiCRISPRv2-GFP

lentiCRISPRv2-mCherry

lenti-sg(MS2)-puro

For any other vector, contact us.

 

Turn around time: 3-4 weeks

Cost: See Product prices (Contact us for large orders)

 

Custom Small Single-stranded Donor Oligo Design

Small single-stranded oligo (ssODN) are used as donor templates for knocking-in point mutations or small tags. Contact us if you need assistance with designing and synthesizing ssODN templates for your genome-engineering needs.

 

Custom Donor Plasmid Design and Cloning

We can design and clone gRNA and donor plasmid/templates to knock-in reporter genes (GFP, RFP, mCherry, puromycin, split GFP, etc.) at the 5'- and or 3'- ends of gene(s) of your interest to create unique engineered cell lines.

 

Custom Pooled CRISPR Panel

A Pooled CRISPR Panel is used to perform a pooled screen (where all gRNA are in a pool, instead of an array). We can design a

pool of CRISPR gRNA to target your specific genes of interest, and clone them into the vector backbone of your choice. If in a lentiviral backbone, we can also make the lentiviral particles for the screening experiment. Contact us for more information.