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GenTarget

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SAN DIEGO, California, US

About GenTarget

Incorporated in 2008, GenTarget provides proprietary bio-reagents and services with an emphasis on Lentivirus and Adenovirus gene delivery technologies, providing Pre-made Lentiviruses and Adenovirus for gene over-expression, knockdown, miRNA expression, Pre-made mammalian stable cell lines, and more.

With... Show more »

Incorporated in 2008, GenTarget provides proprietary bio-reagents and services with an emphasis on Lentivirus and Adenovirus gene delivery technologies, providing Pre-made Lentiviruses and Adenovirus for gene over-expression, knockdown, miRNA expression, Pre-made mammalian stable cell lines, and more.

With an emphasis on product quality and innovation, we develop and manufacture many proprietary technologies such as the SureTiterTM Lentiviral system; LocLightTMLive cell imaging, LoxP GFP/RFP ColorSwtich for CRE reporting system, and EcoTM instant PCR cloning method, as well as wide selection of Signal Transduction Pathway Report Lentivirus product series.

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Our Services (6)


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Lentivirus Production

Price on request

Top quality, Best price and Fastest turnarund time, with >15 year lentivirus experience!

(1) shRNA lentiviral services: 

We construct shRNA target knockdown lentivector by using either your provided shRNA or  our designed shRNA sequences, for your genes of interest. We then generate 0.5 ml of... Show more »

Top quality, Best price and Fastest turnarund time, with >15 year lentivirus experience!

(1) shRNA lentiviral services: 

We construct shRNA target knockdown lentivector by using either your provided shRNA or  our designed shRNA sequences, for your genes of interest. We then generate 0.5 ml of each shRNA lentivirus, and ship you both the shRNA lentivector plasmid DNA and the ready-to-use shRNA lentivirus.

- You choose the shRNA lentivectors 
- We deliver the sequence verified shRNA lentivector and ready-to-use high-titer shRNA lentivirus, including the negative control shRNA (Neg-Control) .


Note: Knockdown validation is not included in this service, but if you order 3 shRNAs for one target, we guarantee either at least one shRNA showing >75% knockdown or a knockdown level of >75% by using the pooled three shRNA lentiviruses.

(2) Over-Expression lentiviral services:  

We construct the target expression lentivector with the features according to your request, and produce the ready-to-use over-expression lentivirus for your desired target.  

- You choose the expression lentivector for the promoter and the marker ;
- We sub-clone your target codon sequence into our lentivector;
- We deliver the high titer, ready-to-use lentivirus to you (note: the sub-cloned lentivector plasmid is not included);


You have the following options with this service:

- Promoters:  suCMV, inducible CMV, EF1a, or CAG
- Selection markers: Blasticidin or Puromycin or antibiotic-fluorescent fusion dual marker
- Fluorescent tag-target fusion
- Any Tags at either the N- or C-terminus
- N- terminus fluorescent fusion marker (GFP, RFP or CFP);
- Or bicistronic expression where target and a fluorescent marker expressed under the same promoter as two separate proteins;

Note: Our optional inducible CMV promoter can be used for either constitutive or inducible expression.

(3) Signal reporter lentivirus services: 

We construct the lentivector by sub-clone your desired promoter into our promoter-less lentivector, and deliver you the ready-to-use lentivirus.  

- You choose the signal reporter type and selection marker;
- We sub-clone your promoter sequence into our lentivectors;
- We deliver the high titer, ready-to-use lentivirus to you 

(4) miRNA expression and anti-miRNA lentivirus services:

- miRNA expression:

We construct an expression lentivector and produce ready-to-use lentivirus for the precursor microRNA expression of any human or mouse miRNA  listed in miRBase database.

- Anti-miRNA: 

We construct an anti-miRNA lentivector and produce ready-to-use lentivirus for inhibition of any miRNA listed in the miRBase database, or any your desired miRNA. You simply select the miRNA you want to inhibit, and we deliver the order within 7-10 business days.

(5) CRISPR / Cas9 lentivirus services:

Gentarget provide pre-made CAS9 expression lentivirus with different antibiotic / fluorescent markers, and design and construct gene specific guild RNA lentivectors (gRNA lentivectors), then produce ready-to-use targeting gRNA lentivirus for gene knockout, and optionally synthesize the single-strand or double-stranded loci specific repair insert or knock-in cassette for gene editing.  See gRNA lentivector core map scheme below. You will receive (1) expression ready humanized CRISPR/Cas9 lentivirus, (2) a set of ready-to-use target gRNNA lentivirus, (3) when requested, the repair / knock-in / mutation / of DNA cassette or single strand repair DNA oligo. 

 

Why use Gentarget’s Lentivirus Services: 

- Take advantage of Gentarget’s extensive hand-on experiences in lentivirus system with many proprietary technologies;
- Use the strongest prompter (suCMV) for the highest protein expression levels in mammalian cells;
- Select most advanced product features for Promoter, Fluorescent tag, Antibiotic selection for  high virus titer;
- Fast turnaround time;
- Best quality and best price in the lentivirus fields;

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Antibody Engineering

Price on request

GenTarget provides antibody engineering and expression services using our own unique optimized protocols. From a hybridoma cell line you provide, we will resolve the variable region and secreted leader sequences of the light- and heavy-chains for isotype determination. We can then insert the variable sequences into our antibody... Show more »

GenTarget provides antibody engineering and expression services using our own unique optimized protocols. From a hybridoma cell line you provide, we will resolve the variable region and secreted leader sequences of the light- and heavy-chains for isotype determination. We can then insert the variable sequences into our antibody expression vectors in either a one- or two-vector system, and assemble them with the  constant regions from human, mouse, or rabbit. The constant regions are codon-optimized for maximum expression in mammalian cells, and the expression leader sequence is engineered for maximum antibody secretion. Upon request, we can also construct any kind of chimeric vector for antibody humanization and engineering.  Please Contact us for a quote for this service.

What we offer:

- Isolate the total RNA and mRNA from collected hybridoma cell pellet;
- Determine the variable and leaser’s sequences for antibody’s light-chain and heavy-chain;
- Clone the determined variable into our mammalian expression vectors;
- Pioneer expression the antibody in suspension 293 cells;
- Stable antibody expression cell line construction;

Why use GenTarget’s antibody cloning / expression service?

- We use a unique protocol for variable region sequence determination
- Our engineered CMV promoter produces extremely high expression levels
- Our engineered leader sequence produces high levels of secreted antibody expression so that the antibody can be purified directly from the media
- The expressed antibody can be humanized for less immunogenicity in human
- Fast turnaround time
- Extensive cloning experience
- The best price
- Upon request, the vector can contain either the glutamine-synthetase gene, the blasticidin resistance gene, or any other marker you select for easier stable cell line construction

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Protein Expression in Mammalian Cells

Mammalian cell based recombinant protein production
Price on request

Mammalian Suspension Expression and Purification: 

Utilizing GenTarget’s advanced lentiviral vector system, we will clone your target into our lentiviral vector with a His tag, generate expression lentiviral particles, and transduce them into mammalian suspension cells for expression. Under... Show more »

Mammalian Suspension Expression and Purification: 

Utilizing GenTarget’s advanced lentiviral vector system, we will clone your target into our lentiviral vector with a His tag, generate expression lentiviral particles, and transduce them into mammalian suspension cells for expression. Under antibiotic selection pressure, only transduced cells will grow and express the target protein. This controlled semi-stable cell line expression system demonstrates high protein expression levels without actually building a stable cell line. The beauty of using the lentiviral system is that lentiviral particles give the highest transduction rate in suspension cells, which can easily generate a large population of target expressing cells. This is a great way to obtain mammalian-expressed protein at a very low cost! 

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Bacterial Protein Expression

Price on request

E Coli Expression and Purification: 

GenTarget’s proprietary Fusion in vivo, PCR cloning technology provides us with an edge when it comes to cloning your target protein into our optimized expression vectors. Our auto-induction medium provides us with a... Show more »

E Coli Expression and Purification: 

GenTarget’s proprietary Fusion in vivo, PCR cloning technology provides us with an edge when it comes to cloning your target protein into our optimized expression vectors. Our auto-induction medium provides us with a worry-free induction protocol for expression. And in case the expressed protein is very toxic, or expressed mainly in an inclusion body, we will use our self-tight-controlled cloning cells. The expressed protein, with a His tag, will be purified by affinity chromatography. 

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Molecular Cloning

Price on request

EcoTM cloning technology 

Our proprietary high-throughput Fusion in vivo cloning technology eliminates the labor-intensive and sometimes troublesome vector preparation procedures of other cloning techniques. It does not need any kind of In Vitro tube reaction, such as... Show more »

EcoTM cloning technology 

Our proprietary high-throughput Fusion in vivo cloning technology eliminates the labor-intensive and sometimes troublesome vector preparation procedures of other cloning techniques. It does not need any kind of In Vitro tube reaction, such as ligation, Topo joining, or In-fusion reaction.

 Let E Coli do the job for you in vivo…and do it directly in the expression strain!

With this technology the same PCR product can be used to make both Gateway-compatible Entry and DEST/expression clones without the use of BP or LR enzymes.

GenTarget’s revolutionary Fusion in vivo high throughput cloning technology (patent pending) is an extremely easy PCR cloning method:

- Simply amplify your gene of interest by PCR  using a primer pair flanked with short arms homologous to the expression vector.
- Next, add 1µl of your PCR product to the engineered, ready-to-use Cloning Cells and
- Immediately proceed to transform the cells.
- You do not need to buy the expression vectors because the expression-optimized E Coli or mammalian expression vector has already been built into the Cloning Cells. (Note: to use your own vector, we can build it into our engineered cloning cells as a custom service). 
For E Coli expression, the verified positive clone will be directly used for T7-based or nonT7-based protein expression (depending on the built-in vector), thus eliminating the second transformation into an expression strain, such as BL21.

Using our Fusion In Vivo technology, We successfully made a large microRNA expression clone collection for one of our clients with a very fast turnaround time and at much lower cost compared to other cloning technologies. We can do the same thing for you! If your collection can be PCRed from genomic DNA or other templates, we can build a collection of shRNA, miRNA or any other.

Advantages of Gentarget’s Fusion In Vivo, EcoTm Cloning Products and Services:

- It is the easiest and most cost-effective PCR cloning method available: simply add 1µl of PCR insert into the provided cells for transformation regardless of the insert’s size and concentration
- No need for the tedious bench work of preparing the vector backbone
- No need for any enzymes or  tube reactions
- Precise directional cloning of PCR products without any extra protein sequences (except affinity tags)
- It does not matter if the PCR products have poly-A overhangs at their 3′-ends. The ploy-A overhang, if there is one, will be removed in the cloning step.
- Compatibility with a wide range different PCR sizes, from 200bp to 10kb
- Engineered E Coli and mammalian expression vectors that produce high protein yields
- Exceptionally adaptable for high throughput cloning 

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Cell Line Generation

Price on request

Mammalian stable cell line creation: 

Generate your desired constitutive or inducible expression stable cell lines at the best price! 

GenTarget’s stable cell line service provides a very cost effective way for you to build a cell line. Using our proprietary lentiviral vectors, we will... Show more »

Mammalian stable cell line creation: 

Generate your desired constitutive or inducible expression stable cell lines at the best price! 

GenTarget’s stable cell line service provides a very cost effective way for you to build a cell line. Using our proprietary lentiviral vectors, we will subclone your target gene into our lentivector, generate the lentivirus particles, sequentially infect the cell line of you request, and select the cell clones that consistently expressing your gene at high levels.

For an inducible expression cell line, we will first infect the cells with TetR repressor-expressing particles to make a TetR stable cell line, then infect with the target expression particles to generate the tetracycline-inducible stable cell line.

Our proprietary suCMV promoter has the strongest expression efficiency available, and will provide extremely high levels of expression for your target. Our unique technology ensures a fast turnaround time. Cell lines are shipped as cryopreserved vials after validating the target gene’s integration. We will work with you to ensure that the cell line meets your requirements so that you will get the best quality and the best price. 

GenTarget Will: 

- Construct a lentiviral vector containing your gene and desired selection marker
- Generate lentivirus
- Transduce the cell line you request
- Select the stably transduced, high-expression cells
- Validate integration of the gene via genomic PCR
- Validate the highest-expressing clone by Western Blot (if applicable)
- Deliver two cryogenic vials of stable cells along with a final report 

What are the advantages of using lentivirus to generate stable cells?

To make stable a cell line, the target has to be integrated into the host cells’ genome for stable, constitutive expression. Random integration (such as by transfection) demonstrates widely varying levels of expression depending on the integration site. Additionally, random integration often results in independent integration of the target and the selection marker, necessitating large-scale screening of select positive clones. Typically less than ten percent of antibiotic-resistant clones express the transgene.  In contrast, lentivirus transduction tends to integrate at high transcription level sites (hot-spots) with a full virus genome. Engineered lentiviral transfer vector that is embedded with a matrix-attachment region (MARs) sequence may provide position-independent transgene expression. Compared to conventional stable cell line construction, transduction with lentivirus produces a much higher positive clone rate and the target always co-exists with the selection marker, therby substantially reducing cost, labor, and time spent selecting stable high expression clones.

Why You Should Use GenTarget’s Stable Cell Line Service: 

- Engineered in-house lentiviral vectors provide high efficiency gene integration with extremely low false positives
- Safe, self-inactivating, high-titer lentiviral particles can deliver your gene into a wide range of cell lines including non-dividing, primary, or stem cells
- Target protein expression can be traced and monitored by fluorescence imaging, allowing sorting or selecting of integrated cells (the fluorescent protein is not fusioned with your target).
- Our experts have years of experience in lentiviral cloning and expression
- Fast turnaround time
- The best price 

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Jack Wang

President

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