MISSION^® shRNA Plus the Control of Targeted Integration

What is the targeted integration shRNA kit?

Some shRNA researchers have found that random lentiviral integration can sometimes interfere with non-targeted genes, causing an undesired phenotype. The MISSION Targeted Integration shRNA kit sets out to eliminate this result. The kit enables researchers to insert their shRNA into the AAVS1 locus in the human genome using our CompoZr zinc finger technology. The AAVS1 locus is a “safe harbor site” that has been validated for safe expression of a transgene^1,2. This allows targeted knockdown of gene expression from a single, defined locus. Targeted insertion of shRNA into a single locus eliminates random lentiviral integration, reducing off-target effects. This kit is produced custom with your specified shRNA pre-cloned into the donor vector, making it ready-to-use with your cell line of choice.

Features and Benefits | Ordering Information | Kit Components | Product Information | Workflow |
Application Data | Vector Map

Features and Benefits

• Validated pair of zinc finger nucleases (ZFNs) engineered to target AAVS1 locus enables efficient targeted integration of shRNA encoding sequence
• Uniform integration into target locus reduces likelihood of expression variability across experiments
• Consistent site for transgene delivery as AAVS1 site is likely present in every human cell line
• No random integration leads to better control of target gene expression
• Effective gene knockdown comparable to lentiviral shRNA constructs

Ordering Information

To inquire about or order our MISSION Targeted Integration shRNA Kit, please contact your local Sigma sales representative or click below and we will put you in touch with a sales rep who will place the order.

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Kit Components

*Custom cloning of any shRNA sequence from our vast MISSION shRNA library from the TRC or choose your own shRNA sequence

Product Information

SKU: CSTPZD
Price: Inquire
Storage: -20° C
Shipping: Dry Ice
Availability: On-demand – ships within 6 weeks
Format: Suitable for nucleofection
Ordering: Contact us at MISSIONRNAi@sial.com or your local Sigma^® Life Science sales representative

Technical Bulletin

Technical Bulletin
(323 Kb PDF)

Workflow

Application Data

Targeted shRNA integration in K562 cells

Figure 1. The indicated shRNA-encoding sequence was delivered to K562 cells with mRNA encoding a pair of ZFN-reagents required for integration of the shRNA-expression construct into the AAVS1 site. Expression of the targeted mRNA following knockdown was measured by qRT-PCR and compared to cells expressing a non-targeting shRNA from the AAVS1 site. Bars represent the average mRNA levels from qRT-PCR samples (n=3). Error bars indicate standard deviation.

AAVS1 shRNA knockdown comparable to lenti-shRNA knockdown

Figure 2. The indicated shRNA-encoding sequence was delivered to DLD-1 or A549 cells by lentiviral transduction or by transfection with ZFNreagents required for integration of the shRNA expression construct into the AAVS1 site. Expression of the targeted mRNA following knockdown was measured by qRT-PCR and compared to cells expressing a non-targeting shRNA. Bars represent the average mRNA levels from qRT-PCR samples (n=3). Error bars indicate standard deviation. All lentiviral data are from pooled populations. AAVS1 data for the first three targets were collected from pooled populations, and data for HIF1A were collected from a clonally isolated cell line.

Vector Map

References

1. DeKelver et al., 2010 Genome Research
2. Lombardo et al., 2011 Nature Methods