© 2018 Functional Genomics Facility

 

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.