|The following is a list of main and recurring characters in the Miss Spider's Sunny Patch Friends series.
- Miss Spider is the mother of the Spider children. She is voiced by Kristin Davis.
- Squirt is a curious young spider who loves web-surfing. He is voiced by Scott Beaudin.
- Shimmer is a strong-willed jewel beetle. She is voiced by Rebecca Brenner.
- Dragon is a bossy and somewhat arrogant dragonfly. He is voiced by Mitchell Eisner.
- Bounce is a fun-loving and adventurous bedbug. He is voiced by Julie Lemieux.
- Holley is the children's music-loving father. He is voiced by Robert Norman Smith.
- Spinner is a bespectacled spider. He is voiced by Austin Di Iulio (season one) and Cameron Ansell (seasons one and two).
- Snowdrop and Pansy are two identical twin spiders. They are voiced by Alexandra Lai and Aaryn Doyle, respectively.
- Wiggle is a shy and self-conscious blue spider. He is voiced by Marc McMulkin.
- Spiderus is the Spider family's villainous neighbor. He is voiced by Tony Jay.
- Ned and Ted are two troublesome ants who love food. They are voiced by Jonathon Wilson and Philip Williams, respectively.
- Stinky is a stinkbug who tries to mask his scent with sweet-smelling flowers. He is voiced by Scott McCord.
- Felix is a frog who refuses to eat bugs. He is voiced by Richard Binsley.