tj@alum.mit.edu
Mentor: Anton McCaffrey, Ph.D.
Lab Phone: 335-7870

Developing Nucleases Specific for Hepatitis B Viral DNA

Although an effective HBV vaccine exists, 400 million people are chronically infected. HBV is the 9th leading cause of death worldwide. Current therapies for HBV are only partially effective, possibly because these therapeutics do not attack existing stores of viral DNA.

We established a proof of principle for a complementary therapeutics, Zinc Finger Nucleases (ZFNs), that specifically target the genomic DNA of hepatitis B virus (HBV). We have designed ZFNs that specifically cleave HBV target DNA in tissue culture and have characterized their specificity and efficacy. Some of the resulting cleaved HBV target plasmids remained linear and some were inactivated by cellular mis-repair.

We developed bioinformatic methods for searching for possible ZFN target sites and possible off-target sites in a number of genomes.



Ramirez CL, Certo MT, Mussolino C, Goodwin MJ, Cradick TJ, McCaffrey AP, Cathomen T, Scharenberg AM, Joung JK. Engineered zinc finger nickases induce homology-directed repair with reduced mutagenic effects. Nucleic Acids Res. 2012 Feb 28. [Epub ahead of print] PubMed PMID: 22373919.

Cradick TJ, Ambrosini G, Iseli C, Bucher P, McCaffrey AP. ZFN-site searches genomes for zinc finger nuclease target sites and off-target sites. BMC Bioinformatics. 2011 May 13;12:152. PubMed PMID: 21569489; PubMed Central PMCID: PMC3113941.

Cradick TJ, Keck K, Bradshaw S, Jamieson AC, McCaffrey AP. Zinc-finger nucleases as a novel therapeutic strategy for targeting hepatitis B virus DNAs. Mol Ther. 2010 May;18(5):947-54. Epub 2010 Feb 16. PubMed PMID: 20160705; PubMed Central PMCID: PMC2890117.