Advances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We test 44 different Cas13 variants to identify enzymes with the best overall performance and show that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also develop Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level.
SummaryAdvances in CRISPR technology have immensely improved our ability to manipulate nucleic acids, and the recent discovery of the RNA-targeting endonuclease Cas13 adds even further functionality. Here, we show that Cas13 works efficiently in Drosophila, both ex vivo and in vivo. We tested 44 different Cas13 variants to identify enzymes with the best overall performance and showed that Cas13 could target endogenous Drosophila transcripts in vivo with high efficiency and specificity. We also developed Cas13 applications to edit mRNAs and target mitochondrial transcripts. Our vector collection represents a versatile tool collection to manipulate gene expression at the post-transcriptional level.
meta-Aminophenols are formed by the action of
DBU on 3-amino-2-chlorocyclohex-2-en-1-ones at room temperature in
MeCN. The chloro compounds are generated by treating 3-aminocyclohex-2-en-1-ones
with the easily prepared halogenating agent BnNMe3·ICl2 in MeOH-CH2Cl2. The amino group must
carry two substituents, either two aryl, one aryl and one alkyl, or
two alkyl groups; 3-aminocyclohex-2-en-1-ones of this type are readily
made from cyclohex-2-en-1-one and a primary or secondary amine.
A multi-well continuous CIDT approach with inline racemization of the solution phase is presented. Using two in-house built PATs and a flow reactor, we were able to successfully crystallize an...
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