RNAitechnology is supposedly next big thing in biotech after recombinant DNA technology.
Fortune has dubbed RNA
interference
biotech''s next "billion dollar breakthrough" in a cover story.
Sciencenamed it the "breakthrough of the year" for 2002
Motivation. CCMB scientist Utpal Bhadra developed drug for cancer treatment using this technology. What is RNA
Interference?It is the mechanism by which double stranded RNA (dsRNA) suppresses the expression of a gene bearing its complementary sequence.Successful implementation of this technology requires the design of double stranded RNA (dsRNA) identical to specific regions of the target gene.
.Protocol includes;1.Choose target starting with AA
2.Av oid stretches of > 4A’s in the target sequence because they act as termination signal for RNA pol III
3.Eliminate from consideration any target sequences with more than 16-17 contiguous base pairs of homology to other coding sequences ( Use BLAST). Some existing software to choose the target sequence on the mRNA.
What problems arise while introducing this method in mammals?In mammalian cells,
introduction of long dsRNA (>30nt) initiates a potent antiviral response due to nonspecific inhibition of protein synthesis and RNA degradation. Remedy:Introduction of double stranded siRNA instead of dsRNA
What are challenges to be encountered?
1.Validating targets 2. Delivery of RNAi in mammalian cells.