Human Genome Project

Chemical Fragmentation

• Maxam and Gilbert

• 1976

• End labeled DNA then chemically modified DNAs creating breaks at specific bases. When fragments were run on a single base resolcing gels DNA seqeunces can be determined. Required specific A, T, G, C reactions - each run on a separate lane.

Chain termination ddNTPs

• Sanger

• 1977

• Utilized fluorescently labeled ddNTPs to terminate and label DNAs during primer extended DNA synthesis. DNA fragments varying in single base size differences were resolved on gels or capillaries. All four ddNTPs could be detected in a single lane or capillary using fluorescence (different emission spectra).

Public

• Francis Collins lead NIH NGP

• Distributed effort across 8 centers.

• Physical map, sequence clone by clone.

• Highest quality possible.

• Public release of data.

Private

• Craig Ventor lead Celera

• Ventor’s lab was funded by NIH, but NIH would not fund his genome sequencing approach.

• Private funding from Pharma in exchange for early acess to sequecnes for drug design (and IP).

• Public funding via stocks ($1 billion peak)

• Shotgun sequence and use computers to assemble gene/unique sequences; ordered using public maps.

HeLa Cell lines

immortal cell lines used worldwide for the study of human disease.

Human ENCODE project

• GOAL: Identify and characterize the non-protein coding regulatory content of the human genome (software)

Results

• The science of comparative genomics - insight into the origin, similarity, and uniqueness of the human genome.

• Accelerated next generation DNA sequencing technology and wide range of discovery in many organisms.

• Diagnostics for inherited traits.

• New drugs targeting specific diseases based on knowledge of genes and encoded proteins identified through positional cloning.

• Gene therapy, gene editing.