Design and cloning strategies for constructing shRNA expression vectors

Glen J McIntyre¹^,2 and Gregory C Fanning¹^,3

¹Johnson and Johnson Research Pty Ltd, 1 Central Ave, Australian Technology Park, Eveleigh, NSW, 1430, Australia

²The school of Biological and Biomolecular Sciences, The University of New South Wales, Sydney, NSW 2052, Australia

³Tibotec BVBA, Gen De Wittelaan L 11 B3, 2800 Mechelen, Belgium

author email corresponding author email

BMC Biotechnology 2006, 6:1doi:10.1186/1472-6750-6-1


Published:	5 January 2006

Additional files

Additional File 1:

A survey of studies that employed expressed shRNA revealed that all shRNA constructs are made from one of three possible methods. A random selection of published studies using expressed shRNA were surveyed and scored for their method of shRNA construction which could be classified as one of three different strategies (see main text for detailed descriptions); (i) Annealed complementary oligonucleotides, (ii) Promoter based PCR or (iii) Primer extension.

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Additional File 2:

Diagrams for shRNA template generation via complementary annealed oligonucleotides or primer extension using Phi29 DNA polymerase. Both the most common shRNA insert construction technique (complementary annealed oligos) and our proposed alternative (primer extension using Phi29) are diagrammatically represented indicating oligo alignments and the features of each oligo.

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