Description
Reverse transcriptase (RT) uses the RNA template to synthesize a complementary DNA strand. It was found in retroviruses. The dCas9-RT fusion was constructed by coupe the dCas9 with the M-MLV RT’s polymerase domain (without the Ribonuclease H domain, i.e RNase H minus). Three desired mutation were made in this RT’s domain for the enhanced Reverse Transcriptase’s fidelity and activity. It can be a powerful genomic sequence repair tool (Reference)
This premade lentivirus express the dCa9-RT fusion under the enhanced CMV promoter (SuCMV), carrying Puromycin selection. The dCas9 introduce a nick in the non-target strand. The RT convert ssRNA (that designed sequence in guild RNA, so called the primer binding sequence) into double-stranded DNA (dsDNA) that is imported into the nucleus and integrates into the host cell genome, leading to DNA modifications or manipulations (Reference).
If desired, the negative control lentivirus, CAT#: LVP1594, which has the dCas9 fusioned with a Null sequence, can be used as the non-specific, background control.
To edit your desired sequence, simply apply this lentivirus to your cells and then add the targeting gRNA lentivirus (which need to be designed and made separately according to your target locus.), or apply this lentivirus together with your desired gRNA lentivirus. (Note: Contact us if you need to make your desired gRNA lentivirus via a customized order).
see details in Product Manual.
Amount: 200 ul/vial, at titer of 1×108 IFU/ml, provided in PBS solution (premixed with 10x polybrene, 60 ug/ml).
CAT#: LVP1566
