Koen J. T. Venken

Contact Information

Research Interests

P[acman], Recombineering to the Fly: Design and Development of a Recombineering Platform to Facilitate In Vivo Structural and Functional Studies of Genes

BAC recombination and modification, also known as recombineering, have been recently introduced into the mouse field. Recombineering utilizes in vivo recombination properties in E. coli to allow gap repair of any desired DNA fragment, allowing the retrieval of DNA fragments while avoiding conventional restriction enzyme cloning. Recombination functions are provided by plasmid borne or genome inserted genes from E. coli or bacteriophage λ. The P[acman] vector was constructed by combining a conditional amplifiable BAC with a P element with transposase sites, a white⁺ dominant marker and an internal multiple cloning site. This provides a BAC transgenic platform for Drosophila to clone any genomic DNA fragment. In addition, by adding an actin promoter or UAS site and eGFP, both separated by FRT or LoxP sites, we provide flox-out or flip-out constructs. These excisions can be catalyzed by heat shock provision of iCre or Flp respectively, to facilitate pre- and postmitotic clonal analysis. Finally, recombineering mediated mutagenesis will allow a rapid in vivo functional analysis of rescue constructs, via P element mediated transformation.

Publications

Venken KJT, Kasprowicz J, Kuenen S, Yan J, Hassan BA, Verstreken P (2008) Recombineering-mediated tagging of Drosophila genomic constructs for in vivo localization and acute protein inactivation. Nucleic Acids Research, Epub ahead of print.

Venken KJT, Bellen HJ (2007) Transgenesis uprgrades for Drosophila melanogaster. Development 134:3571-3584.

Koh TW, Korolchuk VI, Wairkar YP, Jiao W, Evergren E, Pan H, Zhou Y, Venken KJT, Shupliakov O, Robinson IM, O'Kane CJ, Bellen HJ (2007) Eps15 and Dap160 control synaptic vesicle membrane retrieval and synapse development.Journal of Cell Biology 178:309-322.

Venken KJT, He Y, Hoskins RA, Bellen HJ (2006) P[acman]: a BAC transgenic platform for targeted insertion of large DNA fragments in D. melanogaster. Science 314:1747.

Shroyer NF, Wallis D, Venken KJT, Bellen HJ, Zoghbi HY (2005) Gfi1 functions downstream of Math1 to control intestinal secretory cell subtype allocation and differentiation. Genes and Development 19:2412-2417.

Tsuda H, Jafar-Nejad H, Patel AJ, Sun Y, Chen HK, Rose MF, Venken KJ, Botas J, Orr HT, Bellen HJ, Zoghbi HY (2005) The AXH domain of Ataxin-1 mediates neurodegeneration through its interaction with Gfi-1/Senseless proteins. Cell 122:633-644.

Verstreken P, Ly CV, Venken KJ, Koh TW, Zhou Y, Bellen HJ (2005) Synaptic mitochondria are critical for mobilization of reserve pool vesicles at Drosophila neuromuscular junctions. Neuron 47:365-378.

Venken KJT, Bellen HJ (2005) Emerging technologies for gene manipulation in Drosophila melanogaster. Nature Reviews Genetics 6:167-178.

Kazanjian A, Wallis-Schultz D, Au N, Nigam R, Venken KJT, Cagle PT, Dickey BF, Bellen HJ, Gilks CB, Grimes HL (2004) Growth factor independence-1 is expressed in primary human neuroendocrine lung carcinomas and mediates the differentiation of murine pulmonary neuroendocrine cells. Cancer Research 64:6874-6882.

Watase K, Venken KJ, Sun Y, Orr HT, Zoghbi HY (2003) Regional differences of somatic CAG repeat instability do not account for selective neuronal vulnerability in a knock-in mouse model of SCA1. Human Molecular Genetics 12:2789-2795.

Wallis D, Hamblen M, Zhou Y, Venken KJ, Schumacher A, Grimes HL, Zoghbi HY, Orkin SH, Bellen HJ (2003) The zinc finger transcription factor Gfi1, implicated in lymphomagenesis, is required for inner ear hair cell differentiation and survival. Development 130:221-232.

Venken K, Di Maria E, Bellone E, Balestra P, Cassandrini D, Mandich P, De Jonghe P, Timmerman V, Svaren J (2002) Search for mutations in the EGR2 corepressor proteins, NAB1 and NAB2, in human peripheral neuropathies. Neurogenetics 4:37-41.

Irobi J, Nelis E, Meuleman J, Venken K, De Jonghe P, Van Broeckhoven C, Timmerman V (2001) Exclusion of 5 functional candidate genes for distal hereditary motor neuropathy type II (distal HMN II) linked to 12q24.3. Annals of Human Genetics 65:517-529.

Venken K, Meuleman J, Irobi J, Ceuterick C, Martini R, De Jonghe P, Timmerman V (2001) Caspr1/Paranodin/Neurexin IV is most likely not a common disease-causing gene for inherited peripheral neuropathies. Neuroreport 12:2609-2614.