The show features a trifle 8-year-old boy, Diego Márquez (voiced by Jake T. Austin, Brandon "Unkown cognomen" and Matthew Hunter, who helps animals in danger, mainly in the rainforest. His cousin is Dora from Dora the Explorer, as revealed in multiple shows. He's a jaguar companion named Baby Jaguar who assists him on adventures. Thomas Sharkey, who's the voice of Baby Jaguar, is also the singing voice of the kangaroo "Austin" in the Backyardigans. Baby Jaguar also appears in Dora the Explorer but inconsistently; on Go, Diego, Go! he takes a more alive role, which includes being able to speak. It's unknown whether Diego's parents let him keep Baby Jaguar as a pet or Baby Jaguar adopted Diego. Diego's grandfather has a strawberry farm, and he made his first appearance in "Green Iguana Helps Abuelito Plant a New Farm!" It was unknown whether he was Diego's maternal grandfather or paternal grandfather. Diego is also very ticklish, which is revealed in "Diego Saves Baby River Dolphin."
Diego is Dora's older cousin; Dora makes several guest appearances in the series, usually without Boots. Diego's 11-year-old sister, Alicia (voiced by child actress Serena Kerrigan), is a computer whiz and also bilingual; she directs the animal rescue calls that come into the center. She also assists Diego in helping the animals they love. Alicia is responsible and kind-hearted and always kind to her little brother. Diego's parents are described as zoologists; they appear in several episodes, but their names are yet to be given. Dora's dad and Diego's dad are brothers. There has also one other older sister, Daisy (which makes Alicia a middle child), who only appears in "The Bobo's Mother's Day!" with Diego saying that she has abode by college. This also makes Diego the youngest child.
Innermost episodes, Diego hears an animal's cry out for help at his rainforest Animal Rescue Center. With help from his friends, gadgets, and viewers at home, he sets away to rescue the animal.
Other characters include Click (voiced by Rosie Perez), a camera that locates the beast bespeaking help; Rescue carry, a courier bag Diego wears that can transform into any object; and the Bobo Brothers, two troublesome Ateles geoffroyi*. The Bobo Brothers can be stopped by shouting "Freeze, Bobos!" and Diego often encourages viewers to help him stop them by shouting it. While they could be compared to Swiper from Dora, they don't cause trouble for Diego advisedly and apologise after doing so.
There are a few recurring animal friends that are commonly rescued by Diego in their entry but appear later to return the favour and aid Diego out. The basic of these is Linda the Llama, who's appeared prominently in four episodes and made a cameo appearance in "Three Little Condors." In the installment, "Egyptian Camel Adventure" actor Adam Alexi-Malle voices the character "Jamal the Camel". Just like Dora the adventurer, the show teaches preschool-age children the Spanish language. However, the show takes a lesser approach to this and focuses more on the teaching of various animals.
Diego is featured in a one-hour installment titled "The Great Dinosaur Rescue" where he, Alicia, Dora, and Baby Felis onca come back one of these days to save a dinosaur; Nick Jr. has yet to show this episode. However, it was featured on Nick Jr. Australia in August 2007 and has had several re-runs as half-hour segments. Two additional one-hour episodes are in production.
Diego also will be starring in active show called "The Great Jaguar Rescue" where he must save Baby Jaguar. This is also the assumption of an episode of the show that aired on January 15, 2007, and was released on DVD on January 16, 2007.
The live edition of Go, Diego, Go premiered in Jan 2007 starring Richard J. Portela in the character of Diego.
Kelly Ripa, along with her son, guest star as Mommy red wolf and her Pup.
In each episode of Go, Diego, Go! there are several songs and uses of Spanish lexicon.
Whenever a carnivorous predator (anacondas) is featured on the show, its diet isn't observed, unlike while herbivores are shown (such as marmosets). Two notable exceptions to this is when an octopus assisting Diego is explicitly shown and discussed eating live crabs (the crabs, however, are not anthropomorphised in as is way that other creatures typically are in the series) and when a child river dolphin who had tickled Diego in "Diego Saves Baby River Dolphin eats the crabs heading toward a falls. Whenever a featured animal is periled by a predator, the prey is lined as being "afraid" of the predator.
