Generating Lentiviral Vectors for Tumor Suppressor Genes

The SLIEW lentiviral vector was previously cloned [15 (link)] by insertion of a luciferase coding sequence into the BAMH1 site at position 8256, of pHR-SINcPPT-SIEW (SIN-SIEW) [16 (link)]. Consensus coding sequences (CCDS) c-DNA for candidate tumour suppressor genes (IDs - Table S10), synthesised (Life technologies) and cloned into pDONR221, were inserted into the BAMH1 site of a modified SIN-SIEW vector by Gateway^TM cloning (Invitrogen-Fisher Scientific). Inserts of resulting SIN-SIEW-cDNA clones were validated by PCR (Table S10) and Sanger sequencing (DBS genomics, Durham, UK).

Free full text: Click here

Sinclair P.B., Cranston R.E., Raninga P., Cheng J., Hanna R., Hawking Z., Hair S., Ryan S.L., Enshaei A., Nakjang S., Rand V., Blair H.J., Moorman A.V., Heidenreich O, & Harrison C.J. (2023). Disruption to the FOXO-PRDM1 axis resulting from deletions of chromosome 6 in acute lymphoblastic leukaemia. Leukemia, 37(3), 636-649.

Publication 2023

Cdna Clones Coding dna sequences Consensus dna sequences Insertion sequence Luciferase Sequence insertion Tumour suppressor genes Vector

Corresponding Organization : Newcastle University

Other organizations : Teesside University, Princess Máxima Center

Top 5 similar protocols

Variable analysis

independent variables

Insertion of a luciferase coding sequence into the BAMH1 site at position 8256, of pHR-SINcPPT-SIEW (SIN-SIEW) vector
Insertion of consensus coding sequences (CCDS) cDNA for candidate tumour suppressor genes into the BAMH1 site of a modified SIN-SIEW vector by Gateway cloning

dependent variables

Measured outcomes of the candidate tumour suppressor gene cDNA insertions in the modified SIN-SIEW vector

control variables

The original pHR-SINcPPT-SIEW (SIN-SIEW) vector

controls

Positive control: Luciferase coding sequence insertion into the SIN-SIEW vector
Negative control: Not explicitly mentioned

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!