Difference between revisions of "Part:BBa K4830017"
Aruzhan001 (Talk | contribs) |
Aruzhan001 (Talk | contribs) |
||
| Line 5: | Line 5: | ||
Ll.ltrA reverse transcriptase is one of proteins found in The Lactococcus lactis Ll.LtrB intron, that binds the intron RNA to promote RNA splicing and intron mobility. | Ll.ltrA reverse transcriptase is one of proteins found in The Lactococcus lactis Ll.LtrB intron, that binds the intron RNA to promote RNA splicing and intron mobility. | ||
| − | <!-- Add more about the biology of this part here -- | + | <!-- Add more about the biology of this part here --> |
===Usage and Biology=== | ===Usage and Biology=== | ||
| Line 13: | Line 13: | ||
To evaluate the performance of various Ll.ltrA, we cotransfected the RT with pegRNA and ngRNA targeting the RNF and HEK3 genomic loci on HEK293T cells and estimated the editing efficiency using sanger sequencing. Another evaluation test was performed on TLR reporter system at 24hr and 72hr transfection time.The results are plotted as bar graph shown below. Comparison was done against the wild-type PE2 and PE2 with truncated RNAse H region (PE-497). | To evaluate the performance of various Ll.ltrA, we cotransfected the RT with pegRNA and ngRNA targeting the RNF and HEK3 genomic loci on HEK293T cells and estimated the editing efficiency using sanger sequencing. Another evaluation test was performed on TLR reporter system at 24hr and 72hr transfection time.The results are plotted as bar graph shown below. Comparison was done against the wild-type PE2 and PE2 with truncated RNAse H region (PE-497). | ||
| + | |||
| + | <html> | ||
| + | <div class = "middle"> | ||
| + | <img src = "https://static.igem.wiki/teams/4830/wiki/bba-k4830000.png" style = "width:600px;height:400px"> | ||
| + | </div> | ||
| + | </html> | ||
<!-- --> | <!-- --> | ||
Revision as of 12:17, 11 October 2023
Ll.ltrA RT
Ll.ltrA reverse transcriptase is one of proteins found in The Lactococcus lactis Ll.LtrB intron, that binds the intron RNA to promote RNA splicing and intron mobility.
Usage and Biology
Ll.ltrA was one of the Reverse Transcriptases we attempted to use as alternative RT for Prime editing.
Characterization
To evaluate the performance of various Ll.ltrA, we cotransfected the RT with pegRNA and ngRNA targeting the RNF and HEK3 genomic loci on HEK293T cells and estimated the editing efficiency using sanger sequencing. Another evaluation test was performed on TLR reporter system at 24hr and 72hr transfection time.The results are plotted as bar graph shown below. Comparison was done against the wild-type PE2 and PE2 with truncated RNAse H region (PE-497).
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]