Lead Inventors: Martin Chalfie Ph.D., Andrea Calixto



Problem or Unmet Need:

Many important genes are also essential to the survival of the organism, thus a system controlling their expression in real time is need to observe their effects. Current methods to do so all have their draw backs. Inducible promoters such as the tetracycline-responsive (Tet) system can sometime trigger spontaneously and require the use of inducing agents that can cause side-effects by themselves. Gene expression control by RNAi is limited in the consistency of the knockdowns, and creating temperature sensitive strains is a time consuming process that can often alter the function of the gene product of interest. The biggest drawback shared by all of these techniques is that their effectiveness varies wildly from gene to gene. The proposed invention describes an alternative method for controlling gene expression that is more consistent from gene to gene and does not require exogenous agents.



Details of the Invention:

This invention provides a mechanism to control gene expression by combining a cassette built from an intronic sequence the C. elegans mec-2 gene and a temperature sensitive allele the touch sensitivity gene mec-8. Normally, mec-8 functions as a regulation of the intronic cassette. By using a temperature sensitive version of mec-8, one can induce the gene expression of any gene attached to the cassette by changing the temperature. This invention makes available a plasmid containing the mec-2 intron 9 gene and a nematode strain cell line( TU218) expressing the mec-8ts mutation. This method has been demonstrated on rde-1, a necessary component of the RNAi response. Thus allowing for the regulation of the RNAi response.



Applications:

• The technology provides a method for inhibiting expression of a wildtype gene product

• Using temperature to induce gene silencing that does not alter gene function

• Using temperature to regulate the RNAi response, allowing for multiple rounds of knockdown in the same animal

• Potentially construct a library of essential genes controlled in this manner



Advantages:

• More genes can be regulated than with other methods

• Elimination of exogenous inducing agents leads to fewer false positive phenotypes

• Offers repeating rounds of turning gene expression "off" and "on" by temperature changes

• Allows the RNAi response to be switched "off" and "on" and adds the time dimension to knockdown studies




Patent Status: Patent Pending (WO/2006/119356) ~ see links below.



Licensing Status: Available for Licensing and Sponsored Research Support







Other Links:

• USPTO - US20090137046A1
• WIPO - WO/2006/119356

Further Information
Jullian G. Jones
Email: TechTransfer@columbia.edu

File Number: M05-078