Center for Causal Transcriptional Consequences of Human Genetic Variation (CTCHGV)
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Principal
investigator Harvard Medical School Co-investigators
Childrens’ Hospital Massachusetts General
Hospital University of California
at San Diego Information about CTCHGV
· Contact for further information · Internal documents (requires password) |
X-ray
structure of the RecA protein complexed with two partially duplexed single
stranded DNAs (ssDNAs) from Chen et.al. Nature
453:489 (PubMed,
RSCB). RecA plays a key role in homologous
recombination (HR) by enabling ssDNA
templates to be paired against complementary sequences in genomic DNA
during DNA repair. CTCHGV proposes to
make extensive use of HR to alter sites of natural human variation near genes
in order to identify variations that cause changes in gene expression
levels. The starred symbol in the
image above is intended to represent a possibly altered base in a template
ssDNA strand that we might be aiming to incorporate into a genome by
induction of HR through creation of a double stranded break, possibly through
expression of a Zinc
Finger Nuclease designed to cut near the site to be altered. CTCHGV will also explore other means of
altering genomic DNA in human cells, including development of a version of Multiplex Automated
Genome Engineering (MAGE) in human cells.
In MAGE, short ssDNA oligonucleotides harboring variations relative to
the genome are incorporated by a RecA-independent mechanism without the
requirement for inducing double-stranded breaks. |
Last modified: 10/18/2010 7:33 AM by John Aach