Inducible Silencing of RepID in Melanoma

Specific silencing of endogenous RepID was achieved using an inducible shRNA-expressing vector, pSingle-tTS-shRNA (Clontech). shRNAs were inserted into the plasmid using the XhoI and HindIII cloning sites and were delivered into 2451 13T melanoma cells48 (link). Stable clones were selected, and cells conditionally expressing shRNA directed against RepID were induced (or not) with doxycycline for 16 days.

Free full text: Click here

Zhang Y., Huang L., Fu H., Smith O.K., Lin C.M., Utani K., Rao M., Reinhold W.C., Redon C.E., Ryan M., Kim R., You Y., Hanna H., Boisclair Y., Long Q, & Aladjem M.I. (2016). A replicator-specific binding protein essential for site-specific initiation of DNA replication in mammalian cells. Nature Communications, 7, 11748.

Publication 2016

pSingle-tTS-shRNA

Cells Clones Doxycycline Melanoma Plasmid Shrna Vector

Corresponding Organization : Center for Cancer Research

Other organizations : In Silico Biosciences (United States), Cornell University

Top 5 similar protocols

Variable analysis

independent variables

Silencing of endogenous RepID using an inducible shRNA-expressing vector, pSingle-tTS-shRNA (Clontech)

dependent variables

Conditional expression of shRNA directed against RepID in 2451 13T melanoma cells

control variables

Stable clones were selected
Cells conditionally expressing shRNA directed against RepID were induced (or not) with doxycycline for 16 days

controls

Positive control: Cells conditionally expressing shRNA directed against RepID induced with doxycycline for 16 days
Negative control: Cells conditionally expressing shRNA directed against RepID not induced with doxycycline for 16 days

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!