Silencing HIF1A Expression with Lentiviral shRNA

To knock-down HIF1A expression, the pGIPZ lentiviral shRNAmir that targets human HIF1A were purchased from Open Biosystems (Thermo Fisher Scientific, Inc.). We used two different shRNAs against HIF1A: V2LHS_132152 (RHS4430–98513964) (shHIF1A#A) and V2LHS_236x718 (RHS443098513880) (shHIF1A#B). A non-silencing pGIPZ lentiviral shRNAmir was used as the control (RHS4346). The production of lentivirus particles and cells infection was performed as previously described [12 (link)]. To obtain 100% GFP-positive cells, puromycin was added into the medium for an additional 10 days.

Free full text: Click here

Cimmino F., Avitabile M., Lasorsa V.A., Montella A., Pezone L., Cantalupo S., Visconte F., Corrias M.V., Iolascon A, & Capasso M. (2019). HIF-1 transcription activity: HIF1A driven response in normoxia and in hypoxia. BMC Medical Genetics, 20, 37.

Publication 2019

pGIPZ lentiviral shRNAmir

Cells Hif1a Human Infection Lentivirus Puromycin Shrnas

Corresponding Organization : University of Naples Federico II

Other organizations : Ceinge Biotecnologie Avanzate (Italy), Istituto Giannina Gaslini

Top 5 similar protocols

Variable analysis

independent variables

ShRNA targeting HIF1A: V2LHS_132152 (shHIF1A#A) and V2LHS_236x718 (shHIF1A#B)
Non-silencing pGIPZ lentiviral shRNAmir as the control (RHS4346)

dependent variables

HIF1A expression

control variables

Lentivirus particles production and cell infection procedure as previously described [12]
Puromycin selection to obtain 100% GFP-positive cells

controls

Positive control: shRNAs targeting HIF1A (shHIF1A#A and shHIF1A#B)
Negative control: Non-silencing pGIPZ lentiviral shRNAmir (RHS4346)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!