Recombinase-Addressable Data (RAD) Modules

Recombinase-Addressable Data (RAD) Modules

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What is it?

A rewriteable recombinase addressable data (RAD) module that reliably stores digital information within a chromosome. RAD modules use serine integrase and excisionase functions adapted from bacteriophage to invert and restore specific DNA sequences. Briefly, production of integrase alone should set a DNA register sequence flanked by oppositional attB and attP (BP) sites, thereby producing an inverted sequence flanked by attL and attR (LR) sites (State “1”). A second independent transcriptional input drives the simultaneous production of integrase and excisionase and should reset the register sequence to its original orientation and flanking sequences (State “0”).

Our core RAD memory element is capable of passive information storage in the absence of heterologous gene expression for over 100 cell divisions and can be switched repeatedly without performance degradation, as is required to support combinatorial data storage. We also demonstrate how programmed stochasticity in RAD system performance arising from bidirectional recombination can be achieved and tuned by varying the synthesis and degradation rates of recombinase proteins. The serine recombinase functions used here do not require cell-specific cofactors and should be useful in extending computing and control methods to the study and engineering of many biological systems.

 

What can it be used for?

Native or heterologous transcription signals can be recorded via the flipping of DNA into one of two states.  The so-flipped DNA is, by default, a constitutive promoter, that can be used to differentially expression either of two genes (flanking the flipping site).  Other elements can be flipped via changes to the "flipee."

 

Where can I find more information?

Rewritable digital data storage in live cells via engineered control of recombination directionality, Jerome Bonnet, Pakpoom Subsoontorn, and Drew Endy, PNAS June 5, 2012 109 (23) 8884-8889, https://www.pnas.org/content/109/23/8884

 

If you've already received the plate, check out the plate layout. All genes are present on the plate in E. coli glycerol stocks. The plate is labelled "Genetic Memory Collection."

 

Genes

Gene Name Freegenes ID
pBad-Xis/Int Reset Flipper pBad-Xis/Int Reset Flipper BBF10K_000494
RAD Module G8-C1 RAD Module G8-C1 BBF10K_000495
pBad-Int Set Flipper pBad-Int Set Flipper BBF10K_000496

Bionet

The bionet enables open peer-peer exchange of functional biomaterials and associated data. This product may also be available from bionet nodes that are more convenient to you. Here are other bionet nodes who may be willing to provide you this specific product.

Name Contact Country
Kenneth Frimpong ken.frimpong {at} thrivusinstitute {dot} edu {dot} gh Ghana
TING-WEI LIN weitinglin66 {at} gmail {dot} com Taiwan
Jae-Seong Yang jaeseong.yang {at} cragenomica {dot} es Spain
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