Super - thanks again for the help.

Fantastic! Thanks so much for helping out. What would be the best way to move that green bar to align with frame 1, but maintaining the current timing? If I shift the main layer back, it effectively starts the animation about 3 seconds earlier than it should.

Shifting layers on the sequencer may be a good shout actually - I needed a few more seconds at the start so I put all the layers into a folder and shifted that along on the timeline. That may be when my issue first appeared.

You're welcome to check it out - http://www.wilx.co.uk/audio/OffsetIssue.anime.zip, this is the beginning of the project with the offset clicking/selecting issue present on frame zero. I'm not very well acquainted with anime studio, so there's a good chance it's something simple/stupid!

Tricky to describe, but on frame zero (and only frame zero) everywhere I click registers as a click below and to the left of where it should be. So the mouse pointer shows up in the correct place, but if I want to select a point, or a shape, or anything, I have to click somewhere up and to the right...