Kumaran Narayanan

Associate Professor and Deputy Head of School (Research)

PhD (University of Melbourne, Australia)

Email: kumaran.narayanan@monash.edu
Telephone: (+603) 5514 6000, ext: 45807 (DID: 5514 5807)
Room number: 4-8-19 (Building 4, Level 8, No. 19)

1) Recombineering: DNA engineering using homologous recombination

Recombineering permits genetic engineering of DNA using homologous recombination in E. coli. Because homologous recombination is not dependent on the presence of suitably placed restriction enzyme sites like conventional genetic engineering methods, recombineering permits more complex manipulations and is not limited by the size of the DNA. Using this technique, various changes can be added to very large (>100 kb) DNA sequences including i) point mutations, ii) deletions, iii) insertions, and iv) gene fusions to develop specific cell lines and animal models of human disease. My current research is directed to better understand the action of the recombineering enzymes on various DNA substrates and to advance improved applications using this system.

2) E. coli as a vector for gene delivery into mammalian cells

Intracellular bacteria such as Shigella, Yersinia, and Salmonella, have evolved the capability to enter mammalian cells by invasion to establish pathogenicity. My research has adapted a non-pathogenic E. coli strain to express the Y. pseudotuberculosis invasin gene to deliver high molecular weight DNA up to 200 kb in size into mammalian cells by invasion. A vector capable of delivering such large DNA can include complete genes together with their introns, exons, and regulatory regions to permit more accurate expression of a genetic locus. Efforts are ongoing to improve the gene delivery efficiency of this vector into various mammalian cell types.

3) Artificial chromosome research

Gene therapy requires development of improved vectors for long-term retention and accurate expression of transgenes in cells. Currently available vectors can deliver genes efficiently into cells but provide only short bursts of expression before they become silenced by the host cell. To overcome these limitations my research is developing artificial chromosome vectors. Artificial chromosomes will 1) enable long-term retention of the delivered transgenes by existing as independent chromosomes that segregate faithfully to daughter cells during cell division, 2) provide long-term gene expression by avoiding DNA integration into human chromosomes, which can cause transgene silencing, and 3) provide correct levels and duration of expression because they carry complete genes along with their surrounding sequences that function to provide regulation.

Selected Journal Papers

Lee, S.W., Lee, C.W., Bong, C.W., Narayanan, K., and Sim, UE (2015). The dynamics of attached and free-living bacterial population in tropical coastal waters. Marine and Freshwater Research. http://dx.doi.org/10.1071/MF14123

Osahor, A.N., Tan, C.Y., Sim, E.U., Lee, C.W., and Narayanan, K. (2014). Short homologies efficiently generate detectable homologous recombination events. Analytical Biochemistry 462: 26-8. doi: 10.1016/j.ab.2014.05.030. 

Chen, Q., Lee, C.W., Sim, E.U, and Narayanan, K. (2014) Induction of protein expression within E. coli bactofection vector for entry into mammalian cells. Human Gene Therapy Methods 25: 40-7. 

Narayanan, K., Sim, E.U, Lee, C.W., Radu, A. (2013). E. coli bactofection using Lipofectamine. Analytical Biochemistry 439:142-4. 

Chen, Q. and Narayanan, K. (2011). Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays. Analytical Biochemistry, 414: 169-171.

Narayanan, K., and Chen, Q. (2011). Bacterial artificial chromosome mutagenesis using recombineering. J. Biomedicine and Biotechnology, 971296, doi:10.1155/2011/971296

Lee, C.W., Ng, A.Y., Bong, C.W., Narayanan, K, Sim, E.U., Ng, C.C. (2011). Investigating the decay rates of Escherichia coli relative to Vibrio parahemolyticus and Salmonella Typhi in tropical coastal waters. Water Research 45:1561-70.

Sim, E.U., Ang, C.H., Ng, C.C., Lee, C.W., and Narayanan, K. (2010). Differential expression of a subset of ribosomal protein genes in cell lines derived from human nasopharyngeal epithelium. J. Human Genetics 55:118-20.

Lee, C.W., Ng, A.Y., Narayanan, K., Sim, E.U., Ng, C.C. (2009). Isolation and characterization of culturable bacteria from tropical coastal waters. Ciencias Marinas 35: 153–167. 

Narayanan, K., Sim, E.U, Ravin, N.V., and Lee, C.W. (2009). Recombination between double-stranded DNA substrates in vivo. Analytical Biochemistry 387: 139-141. 

Narayanan, K. (2008). Intact recombineering of highly repetitive DNA requires reduced induction of recombination enzymes and improved host viability. Analytical Biochemistry 375: 394-396.  

Ooi, Y.S., Warburton, P.E., Ravin, N.V., and Narayanan, K. (2008). Recombineering linear DNA that replicate stably in E. coli. Plasmid 59: 63-71. 

Narayanan, K., and Warburton, P.E. (2003). DNA modification and functional delivery into human cells using E. coli DH10B. Nucleic Acids Research 31:e51. 

Jamsai, D., Nefedov, M., Narayanan, K., Orford, M., Fucharoen, S., Williamson, R., and Ioannou, P.A. (2003). Insertion of common mutations into the B-globin locus using GET Recombination and an EcoRIendonuclease counter-selection cassette. Journal of Biotechnology 101:1-9.

Narayanan, K., Williamson, R., Zhang, Y., Stewart, A. F., and Ioannou, P.A. (1999). Efficient and precise engineering of a 200 kb -globin human/bacterial artificial chromosome in E. coli DH10B using an inducible homologous recombination system. Gene Therapy 6: 442-447. 

 

Molecular genetics

Book

Bacterial Artificial Chromosomes, 2nd Edition (2015): Methods in Molecular Biology Series, Kumaran Narayanan (Editor), Springer, New York 

Book Chapter

Chen, Q. and Narayanan, K. (2015). Recombineering linear BACs. In Bacterial Artificial Chromosomes, 2nd Edition: Methods in Molecular Biology Series, Kumaran Narayanan (Editor), Springer, New York 

  • Pro Vice Chancellor’s Award for Excellence in Administration, Monash University Malaysia, 2012 
  • Pro Vice Chancellor’s Award for Excellence in Research, Monash University Malaysia, 2011 

PhD Students (Present)

  1. Ms. Liannie Chan
  2. Ms. Eveline
  3. Ms. Liew Pei Sheng
  4. Ms. Alviya Sultana

PhD Students (Completed)

  1. Ms. Chen Qingwen
  2. Ms. Lee Woon Ching
  3. Ms. Felicia Chung
  4. Mr. Erik Hui Yew Woh

MSc Student (Present)

  1. Mr. Ong Poh Tek

Honours Students (Completed)

  1. Mr. Khor Jian Ming
  2. Mr. Ali Mehdi
  3. Ms. Amrita Subramanian
  4. Ms. Bhavini Suraiya
  5. Mr. Kenny Tan Chau Yan  

Adjunct Assistant Professor, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, NY.

Journal reviewer: 

  • Nature Communications 
  • Nucleic Acids Research 
  • Molecular Pharmaceutics 
  • Gene Therapy 
  • International Journal of Nanomedicine 
  • Toxicology and Applied Pharmacology 
  • Journal of Biotechnology 
  • Analytical Biochemistry 
  • Molecular Biotechnology 
  • Journal of Gene Medicine