Sandbox WWC7: Difference between revisions

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==Synthetic Applications==

Recent advances in programmable site-directed DNA-binding proteins (such as CRISPR-[[Cas9]]) have shown incredible potential for medical, agricultural, and scientific applications.<ref>doi:10.1126/science.1231143</ref> As a result, it is not surprising that researchers are attempting to develop similar protein-based tools for programmable RNA binding. Unfortunately, few RNA-binding proteins act in a manner which is predictable enough to facilitate convenient RNA manipulation. PPR proteins hold great promise for this application. As described previously, PPR proteins bind RNA nucleotides in a specific and predictable manner, so researchers are attempting to develop custom-made PPR proteins for use in manipulative gene expression experiments.

Recent advances in programmable site-directed DNA-binding proteins (such as CRISPR-[[Cas9]]) have shown incredible potential for medical, agricultural, and scientific applications.<ref>doi:10.1126/science.1231143</ref> As a result, it is not surprising that researchers are attempting to develop similar protein-based tools for programmable RNA binding. Unfortunately, few RNA-binding proteins act in a manner which is predictable enough to facilitate convenient RNA manipulation. PPR proteins hold great promise for this application. As described previously, PPR proteins bind RNA nucleotides in a specific and predictable manner, so researchers are attempting to develop custom-made PPR proteins for use in manipulative gene expression experiments.<ref name = "engineering">DOI:10.1111/tpj.12377</ref>

===Limitations===

As PPR proteins themselves were only discovered recently, there is still a great deal to be learned ~~about them~~ before PPR design becomes a viable technology. Most importantly, the mechanism of specificity must be completely characterized ~~so that manufactured~~ PPR proteins ~~will be entirely specific~~ and ~~not cause off-target effects~~.

As PPR proteins themselves were only discovered recently, there is still a great deal to be learned before specific PPR design becomes a viable technology. Most importantly, the mechanism of specificity must be completely characterized in order to prevent off target binding. Additionally, no entirely new PPR proteins have been engineered, and design strategies must rely on modifying preexisting proteins.<ref name = "engineering">DOI:10.1111/tpj.12377</ref> Finally, the vast majority of PPR proteins exist in plants and operate only within organelles. The reasons or this fact, both evolutionary and mechanistic, are unknown, but it is likely to complicate the process of using PPR proteins in other organisms and in other locations.<ref name = "engineering">DOI:10.1111/tpj.12377</ref>

==References==

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 ==Synthetic Applications==
-Recent advances in programmable site-directed DNA-binding proteins (such as CRISPR-[[Cas9]]) have shown incredible potential for medical, agricultural, and scientific applications.<ref>doi:10.1126/science.1231143</ref> As a result, it is not surprising that researchers are attempting to develop similar protein-based tools for programmable RNA binding. Unfortunately, few RNA-binding proteins act in a manner which is predictable enough to facilitate convenient RNA manipulation. PPR proteins hold great promise for this application. As described previously, PPR proteins bind RNA nucleotides in a specific and predictable manner, so researchers are attempting to develop custom-made PPR proteins for use in manipulative gene expression experiments.
+Recent advances in programmable site-directed DNA-binding proteins (such as CRISPR-[[Cas9]]) have shown incredible potential for medical, agricultural, and scientific applications.<ref>doi:10.1126/science.1231143</ref> As a result, it is not surprising that researchers are attempting to develop similar protein-based tools for programmable RNA binding. Unfortunately, few RNA-binding proteins act in a manner which is predictable enough to facilitate convenient RNA manipulation. PPR proteins hold great promise for this application. As described previously, PPR proteins bind RNA nucleotides in a specific and predictable manner, so researchers are attempting to develop custom-made PPR proteins for use in manipulative gene expression experiments.<ref name = "engineering">DOI:10.1111/tpj.12377</ref>
 ===Limitations===
-As PPR proteins themselves were only discovered recently, there is still a great deal to be learned about them before PPR design becomes a viable technology. Most importantly, the mechanism of specificity must be completely characterized so that manufactured PPR proteins will be entirely specific and not cause off-target effects.
+As PPR proteins themselves were only discovered recently, there is still a great deal to be learned before specific PPR design becomes a viable technology. Most importantly, the mechanism of specificity must be completely characterized in order to prevent off target binding. Additionally, no entirely new PPR proteins have been engineered, and design strategies must rely on modifying preexisting proteins.<ref name = "engineering">DOI:10.1111/tpj.12377</ref> Finally, the vast majority of PPR proteins exist in plants and operate only within organelles. The reasons or this fact, both evolutionary and mechanistic, are unknown, but it is likely to complicate the process of using PPR proteins in other organisms and in other locations.<ref name = "engineering">DOI:10.1111/tpj.12377</ref>
 ==References==
 {{Reflist}}