Just ran through this in my lab/test environment.
I went about the cutover slightly differently from what is already posted and was able to make the change without causing an iSCSI outage. Also I'm using Jumbo Frames.
Home > Networking
1. Precreate new vDS and configure it for Jumbo Frames.
2. Configure the Uplink Port group with the pNICs you want to use (I chose not to reuse the VSS NICs and had extra NICS available for my new vDS)
3. Configure the iSCSI Port group.
4. Click on the new vDS > Summary > Manage Port groups > Teaming and Failover > configure the new iSCSI port group to use the proper pNIC. You need a one to one relationship to keep in best practice. Also in the later steps, you won't see the new iSCSI VMK show up as an option to associate with the iSCSI adapter unless you're one to one.
At the host > Configuration > Networking > vDS
1. Click Mange Virtual Adapters > Create a new VMK for iSCSI. Have a new iSCSI IP address ready to input. Also note that you'll need to make sure that the new VMK can authenticate to the iSCSI LUNS.
2. Associate the new VMK to the new vDS/Port group you created earlier. Take note of the new VMK number.
3. After clicking OK, go back in and configure the VMK for Jumbo Frames.
At the host > Configuration > Storage Adapter
1. Click on the properties of the iSCSI adapter and click on Network Configuration.
2. Add/associate the new vmk you created with the iSCSI adapter.
3. You'll notice that the path status is not green. Click Close and you'll be prompted to rescan.
4. After the rescan is done, go back into the iSCSI adapter properties and you'll see the new VMK lit up.
5. At this point, you can remove the old VMK.
6. At this point, remove the old iSCSI VSS NIC uplinks and test.
I had RDP sessions open to VMs on the affected iSCSI datastores with the clock opened up and watched the second hand keep ticking while I made the change. No Down time.
Note: I am using the new vDS I created for vmotion and iSCSI since both benefit from Jumbo Frames.
After make the change I notice that I'll have the following benefits if I need to add another host.
Adding each new host via the VSS way.
1. iSCSI - Create a VSS for iSCSI, configure it for Jumbo Frames, create VMK and configure it for Jumbo Frames, Associated the new VMK with the iSCSI storage adapter
2. vMotion - Create a VSS for vmotion, create VMK
Add new hosts via the VDS.
1. iSCSI - Add the host to vCenter, add the pNICs to the uplinks, create new VMKs iSCSI and set for Jumbo Frame, Associate the new VMK with the iSCSI storage adapter.
2. vMotion - Similar to above minus the iSCSI adapter association.
Using the vDS method, I save a few steps creating/configuring new VSSs for each new host.