Part:BBa_K2429077
REST Guide Family Member
Parts K2429077 through K2429081 and parts K2429109 through K2429114 are a family of guide sequences used by either a dCas13a variant or is just a lone guide sequence to target the alternative exon in the mRNA that leads to the production of REST protein in cells. These guides lie downstream of a U6 promoter, which is a constitutive promoter typically used for the production of guide RNA. REST is a transcription factor that prohibits the transcription of oncogenes. When the mature mRNA for REST includes an alternative exon containing a stop codon, the resulting protein (known as REST4) is unable to prevent the transcription of oncogenes and can contribute to tumor formation. Application of our system in cells displaying the incorrect REST4 isoform attempts to correct this splicing error. Our team used multiple guide sequences to "tile" along the pre-mRNA to determine what part of the intron is best to target when trying to control its splicing efficiency. Based on the level of fluorescence associated with each guide, we can compare the results to see whether a specific targeted sequence is integral in the process of splicing, and whether the binding of the RNA binding protein to said sequence changes the splicing efficiency.
The guides shift by a few base pairs so we can make more precise conclusions about what intronic sequences are important for splicing.
Parts K-77 through K-81 lead to guide RNA sequences that serve as a guide for the dCas13a protein.
Parts K-109 through K-114 lead to the production of antisense oligonucleotides (ASOs).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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