Lentiviral Service Request Form

Lentiviral service request form

(Please Fax / Email the completed form to the address above)

Customer contact Information:

You Name:

Institute / Company:

Address:

Phone:

Email:

Service Description:

Gentarget provide service to generate: expression lentivirus, shRNA knockdown lentivirus, promoter specific signaling lentivirus; anti-microRNA lentivirus, or microRNA expression lentivirus. Lentivirus titer is >= 1 x 107 IFU/ml.

· For over-expression, we will sub-clone the customer selected genes into Gentarget’s expression lentivectors, and sequentially produce 0.5 ml expression lentivirus.

· For shRNA knockdown expression, we will design and clone shRNA target sequences into our shRNA lentivectors, and sequentially produce 0.5 ml of each shRNA lentivirus.

· For anti-microRNA, we will construct the anti-miRNA lentivector that expresses the anti-sense to the selected miRNA, and sequentially produce 0.5 ml shRNA lentivirus.

· For signaling lentivirus, we will clone your desired promoter sequence into our lentivector to drive a fluorescent marker or luciferase marker, and sequentially produce 0.5 ml shRNA lentivirus.

· For microRNA expression, we will sub-clone the selected precursor microRNA along with its native flanking sequence (total ~300bp) into our 3UTR microRNA expression lentivector, and sequentially produce 0.5 ml shRNA lentivirus.

· For CRISPR / CAS9 genomic editing lentiviurs: we will provide (1) standalone humanized and neuclear penetrating CAS9 expression lentivirus; (2) design and construct gene specific gRNA lentivector and produce gRNA lentivirus; (3) synthesize the desired repair DNA element, or the knockdown-in marker cassette when requested;

Please provide answers the following questions, so we can give you a specific price quote. You only answer the section that related to your service request.

(A) For shRNA lentivirus (see lentivector's core map scheme below):

1. Can you provide the shRNA target sequence (usually 19-25bp) or do you need GenTarget to design the shRNAs for you (if we design it, please provide us the gene name, better the gene accession number)?

Type in:

If to provide your shRNA sequences, please list your shRNA sequences below:

Type in:

2. Please, select the promoter for express the shRNA sequence:

· Human H1 optional inducible promoter,

· Human U6 constitutive promoter,

3. Please, select shRNA lentivector’s fluorescent reference marker (select only one marker):

· GFP-Puromycin Marker,

· GFP -Blasticidin Marker,

· RFP-Blasticidin Marker,

· RFP -Puromycin Marker,

· Puromycin Marker,

· Blasticidin Marker,

4. Gentarget delivery 0.5ml of each lentivirus as default, if you need more virus, please specify the volume (in ml) below.

Type in:

(B) For over expression lentivirus (see the lentivector map schemes below):

1. Gene name (with specie), or accession number :

Type in:

2. Can you provide the gene template (need to provide 5 ug of template plasmid DNA, in any plasmid backbone), or do you need GenTarget to either synthesize it or obtain it from cDNA collection?

Type in:

3. Please select the promoter (select only one promoter) :

suCMV,

EF1a,

Inducible suCMV,

CAG promoter,

4. Please select tag :

No, do not include His-tag,

Put 6His-tag at N-terminal of the gene,

Put 6His-tag at C-terminal of the gene,

Other tags, please provide the sequence and position

Type in:

5. Please select marker (only select one marker):

Use Blasticidin marker,

Use Neomycin marker,

Use Puromycin marker,

Use GFP-Blasticidin marker,

Use GFP-Puromycin marker,

Use RFP-Blasticidin marker,

Use RFP-Puromycin marker,

Use firefly luciferase marker ,

Make the bicistronic expression target and a fluorescent marker (via 2A element) ,

Any specific requests:

Type in:

6. Is 0.5ml of virus enough, or do you need more (if more, please precise)?

Type in:

(C) For promoter signaling lentivirus (see the lentivector map schemes below):

Your choice promoter name and sequence:

Type in:

Please select marker (only select one marker):

Use Blasticidin marker,

Use Neomycin marker,

Use Puromycin marker,

Use GFP marker,

Use RFP marker,

(D) For anti-microRNA lentivirus (see the lentivector map schemes below):

1. microRNA ID, name (with specie), or matured miRNA sequence :

Type in:

2. Please, select shRNA lentivector’s marker (select only one marker):

· GFP-Puromycin Marker,

· GFP-Blasticidin Marker,

· RFP-Blasticidin Marker,

· RFP-Puromycin Marker,

· Puromycin Marker,

· Blasticidin Marker,

3. Please, select the promoter for express the anti-miRNA sequence:

· Human H1 optional inducible promoter,

· Human U6 constitutive promoter,

3. Is 0.5ml of virus enough, or do you need more (if more virus, please precise)?

Type in:

(E) For precursor microRNA expression lentivirus: (see the lentivector map schemes below):

1. microRNA ID, name (with specie):

Type in:

Human and mouse microRNA precursors and its native context sequences (upstream and downstream flanking genomic sequences) were PCR amplified and cloned into a pLenti-TetCMV(miRNA)-Rsv(Puro) lentivector.

3. Is 0.5ml of virus enough, or do you need more (if more virus, please precise)?

Type in:

(F) CRISPR / Cas9 gene editing lentivirus (see the lentivector map schemes below):

· CAS9 expression lentivectors:

· gRNA lentivectors:

Gentarget provide premade CAS9 expression lentivirus with different antibiotic / fluorescent markers, and design and construct gene specific guild RNA lentivectors, then produce ready-to-use targeting gRNA lentivirus, and optionally synthesize the double stranded loci specific knock-in cassette for gene editing.

Describe service request below:

Lentiviral service request form, Page 7 of 7