Een eerste deel omvat een omschrijving van het kloneren van genomisch DNA. De verschillende vectoren die hiervoor gebruikt kunnen worden, worden besproken. In een volgend hoofdstuk wordt een elegante techniek aangeleerd die in staat is eiwitinteragerende partners op te sporen : de ” two hybrid”...
Next – Generation Sequencing (NGS)_______________________________________________ 26
Verschil tussen Next Generation Sequencing (NGS) en Sanger Sequencing ________________________ 26
Basisprincipe van NGS _________________________________________________________________ 27
Third – generation sequencing (TGS) _______________________________________________ 34
Single molecule real-time (SMRT) sequencing ______________________________________________ 34
Nanopore sequencing _______________________________________________________________ 35
Voorbeelden van High – throughput sequencing toepassingen __________________________ 36
Plaatsspecifieke genoommanipulatie __________________________________________ 37
Knouk – out en knock – in via homologe recombinatie ________________________________ 37
Knock – out _________________________________________________________________________ 37
Knock – in __________________________________________________________________________ 38
Conditionele knock-out via Cre – Lox recombinatie ___________________________________ 39
Cre – Lox recombinatie ________________________________________________________________ 39
conditionele knock-out – Uitschakelen van een gen _______________________________________ 40
Voorbeelden CRISPR – Cas toepassingen ____________________________________________ 47
RNA interference___________________________________________________________ 48
Post – transcriptional gene silencing _______________________________________________ 48
Natuurlijk proces van RNAi _______________________________________________________ 48
RNAi in de biotechnologie _______________________________________________________ 50
Aanmaak van siRNA via in vitro transcriptie ________________________________________________ 50
SIrna vectoren _______________________________________________________________________ 52
Gentherapie op basis van virale vectoren ___________________________________________ 62
Car – T immunotherapie _______________________________________________________________ 63
Genetisch gewijzigde (proef) dieren ___________________________________________ 63
Definities _____________________________________________________________________ 63
Micro-injectie in de bevruchte eicel ________________________________________________ 64
Gebruik van embryonale stamcellen _______________________________________________ 65
Creëren knock – out muis door homologe recombinatie ______________________________________ 65
Kruising voor het creëren van transgene muizen met ES – cellen _______________________________ 66
Conditionele knock – out muizen __________________________________________________ 67
Andere diersoorten _____________________________________________________________ 68
Voorbeelden van genetisch gewijzigde dieren _______________________________________ 68
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