Difference between revisions of "Part:BBa K1777008"
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===Results=== | ===Results=== | ||
Forty-eight hours after transfection with pZW1-sno-ACTB into HEK 293T cells, we use the specific circular exon-4 primers to test the circular RNA. <br> | Forty-eight hours after transfection with pZW1-sno-ACTB into HEK 293T cells, we use the specific circular exon-4 primers to test the circular RNA. <br> | ||
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[[File:acRNA-ACTB.jpg]]<br> | [[File:acRNA-ACTB.jpg]]<br> | ||
| − | Both vector without insert pZW1-sno and plasmid with acRNA pZW1-sno-ACTB appear non-specific electrophoretic band. After sequencing, we found 200bp: circular RNA consists of exon4-exon3-exon4, 500bp: circular RNA onsists of exon4-exon5-exon6-exon2-exon3-exon4 while 700bp: circular RNA consists of exon4-exon5- | + | '''Figure 1. Circular RNA form with acRNA (pZW1-sno-ACTB).''' <br> |
| − | + | HEK-293T cells were seeded into a 24-well plate. Forty-eight hours post-transfection, the total RNAs are extracted. Use circular RNA specific primers (have been BLAST on UCSC-In Silico, proved to be specific) doing PCR analysis, Unexpectedly, we amplify other circular RNAs. Extract non-specific gel and sequence it.Both vector without insert pZW1-sno and plasmid with acRNA pZW1-sno-ACTB appear non-specific electrophoretic band.<br> | |
| − | + | After sequencing, we found 200bp: circular RNA consists of exon4-exon3-exon4, 500bp: circular RNA onsists of exon4-exon5-exon6-exon2-exon3-exon4 while 700bp: circular RNA consists of exon4-exon5-exon3-exon4. | |
| − | + | ||
Revision as of 13:02, 20 September 2015
RNA accelerating cyclization
This RNA can bind to the flanking region of exon4 of hsACTB in HEK293T cells and facilitate the back-splicing mechanism. This part can be used to cyclize certain RNA exon.
Fuction
We design this RNA to promote the cyclization of hsACTB in HEK293T cells.
Results
Forty-eight hours after transfection with pZW1-sno-ACTB into HEK 293T cells, we use the specific circular exon-4 primers to test the circular RNA.
Figure 1. Circular RNA form with acRNA (pZW1-sno-ACTB).
HEK-293T cells were seeded into a 24-well plate. Forty-eight hours post-transfection, the total RNAs are extracted. Use circular RNA specific primers (have been BLAST on UCSC-In Silico, proved to be specific) doing PCR analysis, Unexpectedly, we amplify other circular RNAs. Extract non-specific gel and sequence it.Both vector without insert pZW1-sno and plasmid with acRNA pZW1-sno-ACTB appear non-specific electrophoretic band.
After sequencing, we found 200bp: circular RNA consists of exon4-exon3-exon4, 500bp: circular RNA onsists of exon4-exon5-exon6-exon2-exon3-exon4 while 700bp: circular RNA consists of exon4-exon5-exon3-exon4.