39 research outputs found
Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
RNA interference (RNAi), mediated by short double-stranded RNAs, is a
powerful mechanism for posttranscriptional gene silencing. Sustained
expression of short hairpin RNA (shRNA) can be accomplished in
mammalian cells by viral delivery systems. Using lentiviral constructs,
stable gene silencing is established both in dividing and non-dividing
cells. Targeting one single gene can lead to the development of escape
mutants or may be insufficient to silence redundant pathways.
Therefore, simultaneous targeting of multiple genes may be necessary.
We have generated a lentiviral vector-based system for expression of
multiple shRNAs from a single viral vector, which also encodes an EGFP
reporter protein. We show that knock-down of each single gene from
multiple target vectors is achieved at an efficiency comparable to that
obtained after transduction using single target viral vectors. In this
way, we were able to knock-down several members of the human Rho-family
GTPases in T cells. Double and triple knock-down persisted after
multiple passages of the cells. The ability to inhibit two or more
genes simultaneously from one single expression vector further widens
the application spectrum of RNAi, both in functional studies and
therapeutic strategies
Biomarkers for the stratification of latent Mycobacterium tuberculosis infection in humans.
info:eu-repo/semantics/nonPublishe
Multiple gene knock-down by a single lentiviral vector expressing an array of short hairpin RNAs
RNA interference (RNAi), mediated by short doublestranded RNAs, is a powerful mechanism for posttranscriptional gene silencing. Sustained expression of short hairpin RNA (shRNA) can be accomplished in mammalian cells by viral delivery systems. Using lentiviral constructs, stable gene silencing is established both in dividing and non-dividing cells. Targeting one single gene can lead to the development of escape mutants or may be insufficient to silence redundant pathways. Therefore, simultaneous targeting of multiple genes may be necessary. We have generated a lentiviral vector-based system for expression of multiple shRNAs from a single viral vector, which also encodes an EGFP reporter protein. We show that knock-down of each single gene from multiple target vectors is achieved at an efficiency comparable to that obtained after transduction using single target viral vectors. In this way, we were able to knock-down several members of the human Rho-family GTPases in T cells. Double and triple knock-down persisted after multiple passages of the cells. The ability to inhibit two or more genes simultaneously from one single expression vector further widens the application spectrum of RNAi, both in functional studies and therapeutic strategies.info:eu-repo/semantics/publishe
Optimisation of a flow cytometry assay for the detection of Bordetella pertussis specific memory T cell immune reponses.
info:eu-repo/semantics/nonPublishe
The restricted V-H-gene usage is of prognostic value in B-cell chronic lymphocytic leukemia
info:eu-repo/semantics/publishe
VH-gene usage is of prognostic value in B-cell chronic lymphocytic leukemia
info:eu-repo/semantics/publishe
Association de diffĂ©rents profils fonctionnels de cellules T spĂ©cifiques dâantigĂšnes mycobactĂ©riens Ă la tuberculose active et latente.
info:eu-repo/semantics/nonPublishe
Specific memory B cells to 3 Bordetella pertussis antigens after cellular or acellular vaccine, or after whooping cough during childhood
info:eu-repo/semantics/nonPublishe