This article tests if it is possible to piggyback more than one antisense deoxyoligonucleotide (dO) with one vivo morpholino (VMO). The authors previously developed a piggyback knockdown method that was used to knockdown genes in adult zebrafish. In this article, the authors develop a method to knockdown three genes at one time, and by increasing the concentration of VMO by twofold, could knockdown six genes simultaneously. These multiple gene knockdowns will not only increase the efficiency of the method in whole genome-wide knockdowns but will also be useful to study multifactorial disorders.