Quick Knockin: Targeted Transgenesis Using Rosa26 or Hprt Loci

What are the key features of the Quick Knockin™ technology?

genOway's Quick Knockin™ technology is an approach where we fully control and guarantee the insertion of a transgene by targeting so-called neutral docking sites, also known as permissive loci or house-keeping genes (e.g., Hprt or Rosa26).

This allows us to overcome biased phenotypic observations often seen with random integration techniques, and also produce mouse lines that remain stable for many years, to generate:

  • Reliable reproduction of scientific results, and
  • Comparisons between different transgenic mouse lines created by this technique.

What are the advantages of the Quick Knockin™ technology?

  • Controls and guarantees the transgene expression
  • No tandem-repeats integration
  • Avoids position-effect variegations (PEV) and gene silencing
  • No misregulation of neighboring genes and genetic regions
  • Large fragment insertions
  • Tissue specificity, e.g., gene expressions
  • Time-specific gene expressions
  • Transgene expression in an inducible manner (e.g., FLEx or TET On/Off)
  • Co-expression with a second transgene or reporter gene (IRES system)

 

Reduced development time and costs, thanks to:

  • Ready-to-use targeting vectors
  • Our 10-year track record (see a selection of publications below)
  • An efficient production line to routinely perform such model creation

Published Quick Knockin™ models we have created for our clients

Neuroscience:

van de Beek MC, Dijkstra IM, van Lenthe H, Ofman R, Goldhaber-Pasillas D, Schauer N, Schackmann M, Engelen-Lee JY, Vaz FM, Kulik W, Wanders RJ, Engelen M, Kemp S.
C26:0-Carnitine Is a New Biomarker for X-Linked Adrenoleukodystrophy in Mice and Man.
PLoS One. 2016 Apr 28.

Plucińska K, Crouch B, Koss D, Robinson L, Siebrecht M, Riedel G, Platt B
Knock-in of human BACE1 cleaves murine APP and reiterates Alzheimer-like phenotypes.
J Neurosci. 2014 Aug 6. Keywords: amyloid; circadian; cognition; inflammation; memory; strategy

LeBlanc AC, Ramcharitar J, Afonso V, Hamel E, Bennett DA, Pakavathkumar P, Albrecht S
Caspase-6 activity in the CA1 region of the hippocampus induces age-dependent memory impairment.
Cell Death Differ. 2014 May 21.

Gulbins E, Palmada M, Reichel M, Lüth A, Böhmer C, Amato D, Müller CP, Tischbirek CH, Groemer TW, Tabatabai G, Becker KA, Tripal P, Staedtler S, Ackermann TF, van Brederode J, Alzheimer C, Weller M, Lang UE, Kleuser B, Grassmé H, Kornhuber J.
Acid sphingomyelinase-ceramide system mediates effects of antidepressant drugs.
Nat Med. 2013 Jul;19(7):934-8. doi: 10.1038/nm.3214. Epub 2013 Jun 16.

Ryan D, Koss D, Porcu E, Woodcock H, Robinson L, Platt B, Riedel G.
Spatial learning impairments in PLB1Triple knock-in Alzheimer mice are task-specific and age-dependent.
Cell Mol Life Sci. 2013 Jul.

Mühle C, Huttner HB, Walter S, Reichel M, Canneva F, Lewczuk P, Gulbins E, Kornhuber J.
Characterization of Acid sphingomyelinase activity in human cerebrospinal fluid.
PLoS One. 2013 May 2;8(5):e62912. doi: 10.1371/journal.pone.0062912. Print 2013.

Sarzi E, Angebault C, Seveno M, Gueguen N, Chaix B, Bielicki G, Boddaert N, Mausset-Bonnefont AL, Cazevieille C, Rigau V, Renou JP, Wang J, Delettre C, Brabet P, Puel JL, Hamel CP, Reynier P, Lenaers G.
The human OPA1delTTAG mutation induces premature age-related systemic neurodegeneration in mouse.
Brain. 2012 Dec;135(Pt 12):3599-613. doi: 10.1093/brain/aws303.

Duran J, Tevy MF, Garcia-Rocha M, Calbó J, Milán M, Guinovart JJ.
Deleterious effects of neuronal accumulation of glycogen in flies and mice.
EMBO Mol Med. 2012 Aug;4(8):719-29. doi: 10.1002/emmm.201200241. Epub 2012 May 2.

Stoppelkamp S, Bell HS, Palacios-Filardo J, Shewan DA, Riedel G, Platt B.
In vitro modelling of Alzheimer's disease: degeneration and cell death induced by viral delivery of amyloid and tau.
Exp Neurol. 2011 Jun;229(2):226-37. doi: 10.1016/j.expneurol.2011.01.018. Epub 2011 Feb 2.

Platt B, Drever B, Koss D, Stoppelkamp S, Jyoti A, Plano A, Utan A, Merrick G, Ryan D, Melis V, Wan H, Mingarelli M, Porcu E, Scrocchi L, Welch A, Riedel G.
Abnormal Cognition, Sleep, EEG and Brain Metabolism in a Novel Knock-In Alzheimer Mouse, PLB1.
PLoS One. 2011;6(11):e27068. Epub 2011 Nov 11.

Immunology:

McNeill E, Iqbal AJ, Patel J, White GE, Regan-Komito D, Greaves DR, Channon KM
Contrasting in vitro vs. in vivo effects of a cell membrane-specific CC-chemokine binding protein on macrophage chemotaxis.
J Mol Med (Berl). 2014 Aug 1. Keywords: Chemokine; Inflammation; Macrophage; Therapy; Transgenic

Needham LA, Davidson AH, Bawden LJ, Belfield A, Bone EA, Brotherton DH, Bryant S, Charlton MH, Clark VL, Davies SJ, Donald A, Day FA, Krige D, Legris V, McDermott J, McGovern Y, Owen J, Patel SR, Pintat S, Testar RJ, Wells GM, Moffat D, Drummond AH.
Drug targeting to monocytes and macrophages using esterase-sensitive chemical motifs.
J Pharmacol Exp Ther. 2011 Oct;339(1):132-42. Epub 2011 Jul 21.

Metabolism:

Villarroel-Espíndola F, Maldonado R, Mancilla H, Vander Stelt K, Acuña A, Covarrubias A, López C, Angulo C, Castro MA, Slebe JC, Durán J, García-Rocha M, Guinovart JJ, Concha II.
Muscle glycogen synthase isoform is responsible for testicular glycogen synthesis: Glycogen overproduction induces apoptosis in male germ cells.
J Cell Biochem. 2013 Feb 5. doi: 10.1002/jcb.24507.

López-Soldado I, Zafra D, Duran J, Adrover A, Calbó J, Guinovart JJ.
Liver glycogen reduces food intake and attenuates obesity in a high-fat diet-fed mouse model.
Diabetes. 2015 Mar.

Clarke SE, Kang JX, Ma DW
The iFat1 transgene permits conditional endogenous n-3 PUFA enrichment both in vitro and in vivo.
Transgenic Res. 2014 Mar 13.

Cardiovascular:

Sheen CR, Kuss P, Narisawa S, Yadav MC, Nigro J, Wang W, Chhea TN, Sergienko EA, Kapoor K, Jackson MR, Hoylaerts MF, Pinkerton AB, O'Neill WC, Millán JL.
Pathophysiological role of vascular smooth muscle alkaline phosphatase in medial artery calcification. 
J Bone Miner Res. 2015 May. Keywords: Genetic animal models; Matrix mineralization; Preclinical studies; Therapeutics

Smart N, Risebro CA, Melville AA, Moses K, Schwartz RJ, Chien KR, Riley PR.
Thymosin beta4 induces adult epicardial progenitor mobilization and neovascularization.
Nature. 2007 Jan 11;445(7124):177-82.

Oncology:

Sportoletti P, Varasano E, Rossi R, Bereshchenko O, Cecchini D, Gionfriddo I, Bolli N, Tiacci E, Intermesoli T, Zanghì P, Masciulli A, Martelli MP, Falzetti F, Martelli MF, Falini B.
The human NPM1 mutation A perturbs megakaryopoiesis in a conditional mouse model.
Blood. 2013 Apr 25.

Nephrology:

Rosendahl A, Niemann G, Lange S, Ahadzadeh E, Krebs C, Contrepas A, van Goor H, Wiech T, Bader M, Schwake M, Peters J, Stahl R, Nguyen G, Wenzel UO
Increased expression of (pro)renin receptor does not cause hypertension or cardiac and renal fibrosis in mice.
Lab Invest. 2014 Aug.

Reproduction:

Imudia AN, Wang N, Tanaka Y, White YA, Woods DC, Tilly JL.
Comparative gene expression profiling of adult mouse ovary-derived oogonial stem cells supports a distinct cellular identity.
Fertil Steril. 2013 Nov;100(5):1451-8. Keywords: Oogenesis; Stra8; germ cell; oocyte; oogonial stem cell

Dart DA, Waxman J, Aboagye EO, Bevan CL.
Visualising androgen receptor activity in male and female mice.
PLoS One. 2013 Aug 7;8(8):e71694. doi: 10.1371/journal.pone.0071694. Print 2013.

DNA repair:

van den Born E, Vågbø CB, Songe-Møller L, Leihne V, Lien GF, Leszczynska G, Malkiewicz A, Krokan HE, Kirpekar F, Klungland A, Falnes PØ.
ALKBH8-mediated formation of a novel diastereomeric pair of wobble nucleosides in mammalian tRNA.
Nat Commun. 2011 Feb 1;2:172. doi: 10.1038/ncomms1173.

 

Animal Delivery & Health Status

The animals are housed, bred, and delivered by AAALAC-accredited / PHS-insured professional breeders, under virus- and antibody-free (VAF Elite) conditions.

genOway Licenses Secure Your Discoveries and Research

We are the only company that can provide you with the freedom to operate your created rodent model through a broad range of key technologies. (See: http://www.genoway.com/technologies/overview.htm)

You possess a permanent and royalty-free license for the patented and/or proprietary technologies employed. This guarantees you full rights to use such a model for any and all R&D purposes.

You also retain ownership of the deliverables and can patent the model developed. genOway will have no claim on the results generated using the model we provide for you.